Biomek® 2000. High Density Replication System
Contents
1. LI Figure 22 Initial Configuration for Example Method 2 3 To prompt the operator to change the labware at position B6 with a new sample plate add a system pause by selecting the following Pause System icon Figure 23 System Pause icon 4 Next the System Pause dialog will appear The settings for the System Pause dialog are shown in Figure 24 Biomek 2000 31 HDR User s Guide System Pause Message gi _ E message until canceled message donnes Replace sample at 3 Fe Figure 24 System pause for Manual Labware Change 5 Click OK when done 6 Select the 96 well flat plate from the Labware Bar and click position A3 7 Click on the Begin Loop icon to start the loop for the transfers Specify a loop count of 4 as shown in Figure 25 Click OK when done Begin Loop 4 He LOG ny Gern I RITE Figure 25 Initiating a Loop For Example Method 2 32 Biomek 2000 HDR User s Guide Document 609930 AA 8 Enter the transfers from the source plate at position A3 to each of the destination gels as you did in the previous example Since a new sample plate will be used after the fourth transfer add a sterilization step to the last transfer in the loop Figure2. using da amp 5 using HOAT L5 b n gg bs en 3 3 Hh i I 42 Figure 42 Method Screen For Example Method 3 44 Biomek 2000 HDR User s Guide Document 609930 AA Replenishing Liquid in Reservoirs You may replenish the water or ethanol in the reservoirs by using the purge aspirate and bulk dispense functions Make sure a Wash Tool is installed in the tool rack and the correct solution is in the bulk source container CAUTION Do not use the Wash Tool to replenish the bleach solution as it may damage the tool You can place a Wash Tool in the same tool rack as the HDR Fan with the following considerations The HDR Fan must be positioned in tool rack so that it is located in the slot furthest to the left The Wash Tool then goes into the right most slot The tool rack must be positioned at the far left hand side of the Biomek Worksurface as shown in Figure 43 e No Disposal Accessory or Dual Disposal Unit to left of tool rack Biomek 2000 Worksurface Figure 43 Placing a Wash Tool on a Worksurface Because the fan extends over the edge of the worksurface you may receive the warning message shown in Figure 44 when placing the labware Biomek 2000 45 HDR User s Guide Windows Applicat
3. 59 Items Not Covered by 60 OSS ANY Or Ter atu uU 61 ii Biomek 2000 HDR User s Guide Document 609930 AA Notice This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment This equipment generates uses and can radiate radio frequency energy and if not installed and used accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area 15 likely to cause harmful interference in which case the user will be required to correct the interference at his own expense The High Density Replicating System using BioWorks software 16 copyrighted and all rights are reserved by Beckman Instruments Inc Duplicating other than as specified herein selling or otherwise distributing this product is a violation of the law This manual is copyrighted and all rights are reserved This document may not in whole or in part be copied photocopied reproduced translated or reduced to any electronic medium or machine readable form without prior consent in writing from Beckman Instruments Inc The following trademarks are used in this ma
4. High Density Replication Transfer HORT Ey nm Ej zed bart E Directa Decta m ue Figure 19 HDR Transfer for Example Method 1 4 Repeat this process to define the transfer from A5 to the second gel at position B4 When the transfer dialog is displayed check Sterilize to enable a sterilization step to clean the tool before it accesses the next sample plate Use the parameters shown in Figure 20 Biomek 2000 29 HDR User s Guide High Density Replication HDA Transfer Source Labware Location 55 Pause 1 E rection T Deshnation Labware Location 4 Label HDR GEL 2 2 GEL 2 2 Pause 1 sec Tool HDRT Bleach Pause 1055 sec Wi ater Pause 1055 sec Pause 10 Pause 2025 st abel S6 well Hate E ret Hate Sterilize Cancel Help Figure 20 Transfer with Sterilization Step 5 Continue defining transfers from each of the sample plates to the gels in the same manner To achieve the matrix pattern shown in Figure 17 select source plates in the order 5 A6 B5 B6 Use a sterilization step after the second transfer from each plate Wh
5. 9 Now input the loop which will control moving the gels from the Stack Area to the worksurface Click on the Begin Loop icon select Loop for count and specify 5 iterations From the Labware Bar click on the Next Labware icon shown in Figure 35 Click on worksurface position A4 Figure 35 Next Labware icon The Next Labware dialog will appear as shown in Figure 36 Specify Remove current item get next item uncheck Recover lid before Removal then click OK when done Labware Tip Hack Movement Remove current item get next item C Remove current item only lid on nest item Cancel Help Figure 36 Next Labware dialog 10 Repeat this process to add a Next Labware command for positions A5 B4 and B5 11 Go to the Labware Bar and select the Set Shelf icon 12 Click the first shelf of stack A then click on the worksurface position B6 13 The Set Shelf dialog will appear Click OK when done The Method and Sideloader screens as well as the Worksurface screen should appear as shown in Figure 37 Biomek 2000 39 HDR User s Guide xima aa vie aj M EU Se cK 2 Initial Confiquestian irt Shel ICi to Worksource Sket 100 1a 5 161 to Worksurlece Eid Figure 37 Next Labware Commands 14 Within t
6. 26 High Density Replication HDH Transfer m ITem Figure 26 HDR Transfer with Sterilization step for Example Method 2 9 Click OK when done Add the End Loop function Your method will look like the one shown in Figure 27 Biomek 2000 33 HDR User s Guide IFieplicale te Ad ucing uziag HOAT lir pire is dleplicate 21 je B5 HORT sterilize Doom eho ae Figure 27 Method Screen for Example Method 2 34 Biomek 2000 HDR User s Guide Document 609930 AA Example Method 3 HDR Transfer with Side Loader To fully automate the labware placement operations and increase throughput you can employ a Side Loader The following method demonstrates how to use a Side Loader and one Stack Area to automate the handling of labware for an HDR transfer This method has the following characteristics 5 different sets of 4 plates are produced 4 duplicate gel plates each using a 4 x 4 grid pattern are made at a time a total of 20 gels are produced 16 sample plates are used as source labware for the transfers to each of the gels These source plates are moved individually from the stack area to the Biomek worksurface and back by the Side Loader Before beginning this method create a custom Gel with a 4 x 4 matrix see Figure 28 Call the Gel HDR GEL 4 x 4 Refer to Creating a Custom
7. Depth enter the distance from the top of the labware down to the top of the Gel where you wish the HDRT to transfer liquid Click on OK to enter the parameters for the HDR Gel If you wish to create several HDR Gels click on the HDR Gel and then on Copy The Gel will be copied You may enter different parameters and name the copy as you wish it to be listed in the library Creating an HDR Method Note that the following example 15 for a 96 pin transfer For a 384 Pin tool transfer use the same procedure substituting appropriate matrix dimensions and grid spacing values so that the defined matrix pattern does not occupy an area greater than 4 5 x 4 5 mm BioWorks also supports 384 Well plates HDR methods are created in essentially the same way as other methods you set up the initial configuration including sterilization components then specify transfers Loops are often useful for HDR methods because of the multiple transfers made to a single gel For example a 4 x 4 matrix requires 16 transfers from sample plates to the gel By using loops to handle repetitive tasks methods can be simplified and shortened The Biomek automatically advances the deposit positions of the pins for subsequent HDR transfers to the same gel 20 Biomek 2000 HDR User s Guide Document 609930 AA CAUTION The method must specify the correct number of transfers necessary to complete the matrix If the method specifies more transfers than the matrix
8. Guide 2 Select the HDR Gel labware type from the list then click the Copy button The Copy Labware screen will appear From HDR Gea 0 HDR Gel or Membrane Em Mame HDRGEL 2x2 Author Beckman Instruments HDRGel 0 Figure 13 screen Copy Labware On the Copy Labware screen shown in Figure 13 enter a new name such as HDR Gel 2 x 2 then click OK The Edit Labware screen will re appear as shown in Figure 14 24 Biomek 2000 HDR User s Guide Document 609930 AA Edit Labware Author keyword Filter Beckman Instruments HDR Library Labware C Modular Reservoirs Labware MH ame GEL 2 2 Gel or Membrane J6 well v bottom J6 well vinyl Hat Sb well u bat Sb well vinyl v bat Beckman Coming 12 Costar 24 Cryo v bat Falcon 24 Filter Plate FlashPlate Full Reservoir Bleach Ethanol GEL 2 x 2 Gel 384 Gel Vater Labware Type Titerplate or similar Titerplate or similar Titerplate ar similar Titerplate or similar Titerplate ar similar Titerplate or similar Titerplate ar similar Titerplate or similar Tube Module small Titerplate or similar Titerplate ar similar Titerplate or similar Titerplate or similar Titerplate or similar Titerplat
9. OG PPS od ino Hp 5 el Boe r nrc 5 5 Pre Installation 6 Pre Installation 6 6 EIS CUI all 6 a eee 6 Additional Requirements 7 Setting up the HDR System xoci eon erbe i eo 8 eT dus 8 HDR Sterilizer Components xe e olx vo eua 9 eMe NN 9 Water HeSORVOIE ereietuoe Erba aUe vou etos 9 minzipelalae g wm ec ren ea ee 9 FAN CHOI fee 9 High Density Replicating Tool 10 Gable CMDS cs c 10 MISC c 10 Assembly of the HDR Tool 10 Biomek 2000 i High Density Replication System User s Guide Document 609930 AA Installing the HDR Sterilizer 12 Activating the HDR System 8 17 Using the BioWorks HDR Options 17 UO G
10. for additional warning WARNING If you use the tool rack to hold the bleach reservoir remove the tool from the bleach and rinse thoroughly after each ses sion Do not store the HDRT with the pins immersed in bleach when not in use Extended contact with bleach will result in corrosion of the metal pins Cable Clips Beckman Instruments also provides adhesive clips to assist in routing the fan cable or the power supply cable The Gels A Gel is the target to which you transfer your samples See Setting Up HDR Gels The Gel may be a gel membrane or other surface to which submicroliter volumes of liquid are to be transferred The Gel 1s divided into 96 locations for 96 well plates or 384 locations for 384 well plates You specify the number of rows and columns in each location through the software Assembly of the HDR Tool Components To assemble the pieces of the HDR Tool use the following procedure 1 Ifthe pin plate is not attached to the tool body make sure that the pin retainer is securely in place 2 Turn the tool body upside down and attach the pin plate to the bottom with the two screws provided 4 40 x 1 2 flat head stainless steel See Figure 2 10 Biomek 2000 HDR User s Guide Document 609930 AA Mounting Screws cL006 Pin Retainer Tool Body Figure 2 HDR Tool Assembly 3 Loosen the two captive thumbscrews and remove the pin retainer from the tool Store for future
11. of the pin plate 5 For storage turn this assembly plate pins and retainer upside down so it rests on the retainer pins pointing up 4 Pin Plate Mounting Hole HDR Reservoir or 96 well Plate Figure 53 Inserting Pins in Pin Plate of HDRT 56 Biomek 2000 HDR User s Guide Document 609930 AA Sterilization WARNING Beckman does not endorse or certify any specific method of decontamination and or sterilization of biologically active agents Follow the procedures as specified by your Labora tory Safety Officer The following information is given to ensure that the Biomek is not damaged by such decontami nation and or sterilization procedures The pins and the retainer plate of the HDRT can be autoclaved at 121 C for one hour After autoclaving allow the pins to cool to room temperature before handling them All other components should be disinfected with 75 ethanol and or exposure to ultraviolet light In the event of biological contamination follow the appropriate decontamination procedures for your facility Biomek 2000 57 HDR User s Guide Supply List 96 HDR System 120V 50 60 7 267616 Includes a Tool Body 96 Pin Plate Kit and Fan Kit for 120V 50 60Hz operation 96 HDR System 240V 50 60 27 267617 Includes a Tool Body 96 Pin Plate Kit and Fan Kit for 240V 50 60Hz operation 384 HDR System 12
12. one Fan Kit Refer to the Supplies List later 1n this manual HDR Tool Body The HDR Tool Body provides a Biomek compatible tool body which can accomodate either a 96 Pin or a 384 Pin Plate This allows you to use the Plate Kit which best suits your needs and to change from one configuration to the other with the same tool body This kit includes the mounting screws required for attachment of a Plate Kit HDR 384 Pin Plate Kit The HDR 384 Pin Plate Kit includes 2 plates 400 0 015 diameter pins and 400 0 045 diameter pins HDR 96 Pin Plate Kit The HDR 96 Pin Plate Kit includes 2 plates 100 0 015 diameter pins and 100 0 045 diameter pins HDR Fan Kits Each 96 384 Fan Kit includes a fan and power supply 120V or 240V 3 reservoirs 2 reservoir supports a tool rack and 5 cable clips One Fan Kit 1s required for an HDR System HDR Pin Kits Each Pin Kit provides 20 replacement pins There are two pin kits available one with 0 015 diameter pins and one with 0 045 diameter pins Biomek 2000 5 HDR User s Guide Pre Installation Requirements Pre Installation Equipment The HDR System is an accessory to the Biomek 2000 system Before it can be installed be sure that you have the following items in your laboratory e Beckman Biomek 2000 Laboratory Automation Workstation installed as directed in the Installation and Maintenance Guide e BioWorks software version 3 0 or later e Computer with at leas
13. one shown in Figure 40 Biomek 2000 41 HDR User s Guide Lt I 1 pa lume peni rr M Bap LIES nisal Erant Shel 4 Shh do Warksurisre 55 Set Biel Tel Sheli IFI In rplicate ta Ad using eglicate da AS using HOAT leplicese BG using HORT va using HORT Eieriize Himmat ZDUI RA Figure 40 Specifying End Loops for Example Method 3 42 Biomek 2000 HDR User s Guide Document 609930 AA Labware Tip Hack Movement C Remove curent item get next item Remove current item only lid Cancel Help Figure 41 Specifying Labware Removal 19 At the end of the method you should also remove the HDR Gels that were placed on the Worksurface and put them back on the Side Loader stack area To do this highlight the END command in the method then select the Next Labware icon Click on position A4 on the Worksurface then select Remove current item only from the dialog 20 Repeat this for positions A5 B4 and B5 Your method should look like the one shown in Figure 42 Biomek 2000 43 HDR User s Guide 1
14. specifies deposits on the matrix may be contaminated by mul tiple transfers NOTE If the bleach reservoir is in the tool rack then the Biomek will per form a sterilize function automatically before performing the trans fer First double click on the Edit icon in the BioWorks program group Then select Method New Click on Clear Configuration Setting up the Configuration for an HDR Method If you will be using sterilization set up the sterilization components just as you would set up any other labware The bleach water and ethanol reservoirs are under the Plates option on the Labware bar To set up the fan the HDRT and the optional bleach tray in tool racks on the worksurface click on Other on the labware bar and select a tool rack Click on the location where you want to place the tool rack A window similar to the one in Figure 10 appears Tool Change Tool List Tool Rack 2 4 4 Bleach Cancel Help Figure 10 Specifying the Tool Rack Contents Biomek 2000 21 HDR User s Guide NOTE Note that two tool racks are required for the fan and the HDRT You Should use two adjacent locations such as Al and A2 or A2 and A3 for the tool racks If you have a Side Loader or Tip Disposal enabled also be aware of any other restrictions which apply to labware place ment We recommend location Al or A2 for the fan for ease of wir ing See Figure for example Biomek
15. the location of the sterilization liquids using the BioWorks software so that when a sterilization occurs in the method the Biomek will perform the sterilization at the correct location and sequence Each of the components for sterilization is described below Bleach Reservoir Bleach 15 used to sterilize the HDRT pins The bleach reservoir may rest in the same tool rack as the HDRT You may also place the bleach reservoir at any location on the worksurface Warning Remove the Bleach immediately after the method is com plete Water Reservoir The water reservoir is used to remove the bleach residue from the pins of the tool Ethanol Reservoir The ethanol reservoir is used to further sterilize and dry the pins of the tool Fan Unit The fan 15 used to dry the pins of the HDRT after the tool has been immersed in one or more of the sterilization solutions The fan is installed in a tool rack at any appropriate location on the work surface The fan may occupy an entire tool rack or may share the rack with one pipetting or wash tool Biomek 2000 9 HDR User s Guide High Density Replicating Tool HDRT HDRT 15 the 96 or 384 pin tool that transfers the sample to the specified locations on the Gel The HDRT may reside in any tool rack on the work surface You may also place a bleach reservoir under the tool so that the pins are immersed in bleach when the tool is at rest Refer to the Bleach Reservoir section
16. 0V 50 60 7 267618 Includes a Tool Body 394 Plate Kit and Fan Kit for 120V 50 60Hz operation 384HDR System 240V 50 60 2 267619 Includes a Tool Body 364 Plate Kit and Fan Kit for 240V 50 60Hz operation HDR Tool Body 609668 354 Pint Plates 325669174 300 AURA CS RD edi pU e 609079 Includes 2 plates one with 0 015 pins one with 0 045 pins HDR 90 Pin Plates ns aereo e e QR a ae T IP 609004 Includes 2 plates one with 0 015 pins one with 0 045 pins 120 V oo doceo todo Eon pir bus aon aan Gad eee Gon ele fae 609002 Includes 3 HDR reservoirs fan tool rack and power supply 609003 Includes 3 HDR reservoirs fan tool rack and power supply Pri Kit 0015S Meee ee tess 609889 Includes 20 pins Pant 0 045 asd aoe nae 609855 Includes 20 pins 58 Biomek 2000 HDR User s Guide Document 609930 AA Warranty Subject to the exceptions and upon the conditions stated below Beckman warrants that the Biomek 2000 with the High Density Replicating System shall be free from defects in workmanship and materials for one year after delivery of the product to the original Buyer by Beckman and if the pro
17. 2000 Worksurface Figure 11 Typical HDR Setup NOTE Do not place a tool rack of 5 4 137 mm length immedialety to the left of an automatic latch tip rack holder You will need to purchase a new tool holder for this purpose You can also place a wash tool in a tool rack to replenish the water and ethanol Click on the appropriate tool options then click on the desired location in the tool rack If you are using a bleach reservoir under the HDRT enable the Bleach Tray check box NOTE When a bleach reservoir is used under the tool rack the HDRT is automatically sterilized before the first transfer The HDR Ethanol Tray HDR Water Tray and HDR Fan must be present in the method configuration If a sterilization has been specified for the first transfer the pause times for the sterilization will be taken from that sterilization If not the default pause times will be 0 for bleach 10 for water and etha nol and 20 for the fan 22 Biomek 2000 HDR User s Guide Document 609930 AA Use the Plates selection on the labware bar to select the HDR labware items for placement on the worksurface A typical setup may look like the setup shown in Figure 11 If you are using a 384 Pin tool you can select 384 well plate from BioWorks The remainder of this section provides step by step instructions for customizing gel labware and creating some typical HDR methods Creating an HDR Method Examples The next two examp
18. Beckman Part No 609930 AA August 1998 7 High Density Replication System User s Guide BECKMAN Beckman Instruments Inc 4300 N Harbor Blvd Fullerton CA 92835 Copyright 1998 Beckman Instruments Inc Printed in U S A Copyright Licenses and Trademarks Copyright Beckman Instruments Inc 1998 All rights reserved No part of this publication may be reproduced transcribed transmitted or translated into any language in any form by any means without the written permission of Beckman Instruments Inc The product warranty does not apply to software which is warranted on an as is basis without fitness for any specific purpose with the exception of the media which is warranted against defects in materials and workmanship for a period of one year The software is copyrighted and may not be altered or given to a third party without the written authorization from Beckman Instruments High Density Replication System User s Guide Document 609930 AA Table of Contents T 1 NOICE 2 NETTO 4 General DESCTIDTION cccccccsseeccceeeeeeeeeeeceeueecseuseesseueeesseneeessaaeessaaeees 4 HDR System Components 5 FID 5 2 iu i 5 HDR
19. Gel for detailed instructions on creating custom labware E Columns Grid Spacing Columns 124 mm TT TTT Te TT TT YT Well Depth 11 752 Height 26 00 mm Can Have Lid Parameters Lid Length Lid width A Width Lid on Height C Labware End View Lip Height D Acceleration 1 00 iE Grip Type Below bottom Lifter Height Grip at top Height 000 E mm Grip Lid Depth GT Figure 28 4x4 Gel for Example Method 3 Biomek 2000 35 HDR User s Guide 1 The initial configuration should look like the one shown in Figure 29 Note that the sample plates and gels will be placed on the worksurface automatically by the Biomek Biomek 2000 Worksurface Side Loader Top View Setup Port Valve Figure 29 Initial Configuration for Example Method 3 2 Designate each stack in Stack Area as 8 Shelf Stack 3 Now populate the stacks with the labware for the method To populate the first two stacks select the 96 Well Flat plate from the Labware Bar then click on Shelf 1 through Shelf 8 in the first stack Then click on Shelf 1 through Shelf 8 of the second stack Figure 30 Side Loader Stack Area 1 gb wellHat db wellHat 8 Sb well flat SBb well flat SBb well Hat db wellHat 8 96 well flat SE wellHat JE well flat 3 SE wellflat SE vve
20. UICU OV Pr TR 17 Setting up HDR Gel 18 Creating an HDR sisi dta ende aia 20 setting up the Configuration for an HDR Method 21 Creating an HDR Method 23 Creating a Custom Poeta tac 23 Example Method 1 Basic HDR 27 Example Method 2 HDR Transfers within Loops 31 Example Method 3 HDR Transfer with Side Loader 35 Replenishing Liquid in 45 Preparing 10 RUN a 50 RUNNING An HDR 50 Locating opece 51 Troubleshooting amp 52 General Maintenance Procedures 53 53 FAN Fev 53 Replacing the Pin Plate in the HDR 54 Replacing Pins in Pin 56 OLS NIZA Iano aeneae 57 leac 58 m
21. ch 3 to 10 ethanol 95 ethanol and sterile water The recommended volume for liquids used in the HDR reservoirs 15 about 80 mL Biomek 2000 7 HDR User s Guide Setting up the HDR System Introduction A typical HDR System uses both a sterilization system and the High Density Replicating Tool HDRT The sterilization system consists of reservoirs for bleach water and or ethanol and the fan The HDRT which consists of the Tool Body a Pin Plate and appropriate pins resides in a tool rack Additional labware is defined as Gels holds membranes gels or any other receiving medium Figure 1 shows typical HDR Setup HDRT components are described in the following section 900474 Al Figure 1 Typical HDR Setup 3 Ethanol Reservoir 7 Bleach Reservoir 4 Sample Plate 8 Tool Rack 8 Biomek 2000 HDR User s Guide Document 609930 AA NOTE The configuration shown in Figure is not compatible with use of a Tip Disposal HDR Sterilizer Components Each HDR Fan Kit provides three reservoirs for water bleach or ethanol that may be used interchangeably and be placed at any available labware location on the work surface Once sterilized with one or more of these solutions the pins of the HDRT are dried using the fan Typically the sterilization process immerses the pins of the HDRT in bleach then water then ethanol then finally places the tool onto the fan to dry the pins You program
22. cing between deposit positions edit the Gel labware type in the labware library See Setting Up HDR Gel Definitions as follows When an HDR Transfer Function is specified pause times are entered for the worksurface locations where the tool will pick up sample deliver sample and sterilize its pins The rate at which the HDRT approaches the source and Gel is also specified in the Transfer Function Biomek 2000 17 HDR User s Guide Setting up HDR Gel Definitions HDR Gels are the targets for the HDR transfers The default definition is for a Nunc Omni Tray which may be filled with a Gel or covered with a membrane Each Gel is divided into 96 or 384 grid locations which all receive transfers at the same time using the HDRT 96 grid locations are 9mm x 9mm while 384 format grid locations are 4 5mm x 4 5mm The same matrix pattern defined in BioWorks is applied to all grid locations on the Gel The matrix defines the number of deposit positions within each grid location the spacing between positions and the height of the HDRT deposits To define the Gel type for your application select Edit Labware from the menu bar in the Edit module A list of labware is displayed Figure Edit Labware Author Filter Beckman Instrumente HDR Gel Library Labware Last Modified C Modular Reservoirs 06 1 9 98 Labware Mame HDA Gel HDR Gel or Membrane Copy Labware Type viny
23. clean the system However you can autoclave the stainless steel pins and plate and use organic solvents to clean them e Liquid transfers may generate aerosols Therefore operate the Biomek 2000 in an appropriate safety enclosure and take all necessary precautions when using pathologic toxic or radioactive materials e Objects dropped onto the labware or accidental tool release may result in splashing of liquids therefore take appropriate safety precautions such as the use of safety glasses when working with potentially hazardous liquids e Never attempt to exchange labware reagents or tools while the Biomek 15 operating as indicated by the amber warning light on the front of the bridge When the warning light is ON it indicates that the workstation is in operation and may move suddenly and rapidly at any moment e Because the parts of the Biomek move automatically keep clear of the head assembly during operation Also keep the area around the workstation clear including the side module areas to prevent obstructing the movement of the instrument 2 Biomek 2000 HDR User s Guide Document 609930 AA An emergency stop switch is located at the base of the workstation front center in case an emergency stop 15 required Pressing this switch will freeze the workstation instantly stopping the run in progress e When powering up system components NOT specifically described this manual on the accessory power stri
24. duct should prove defective within such one year period Beckman agrees at its option either 1 to correct by repair or at Beckman s election by replacement with equivalent product any such defective product provided that investigation and factory inspection discloses that such defect developed under normal and proper use or 1 to refund the purchase price The exceptions and conditions mentioned above are as follows a b c d Components or accessories manufactured by Beckman which by their nature are not intended to and will not function for one year are warranted only to give reasonable service for a reasonable time what constitutes reasonable time and reasonable service shall be determined solely by Beckman A complete list of such components and accessories is maintained at the factory Beckman makes no warranty with respect to components or accessories not manufactured by it in the event of defect 1n any such component or accessory Beckman will give reasonable assistance to Buyer in obtaining from the respective manufacturer whatever adjustment is authorized by the manufacturer s own warranty Any product claimed to be defective must 1f required by Beckman be returned to the factory transportation charges prepaid and will be returned to Buyer with transportation charges collect unless the product is found to be defective in which case Beckman will pay all transportation charges Beckman shall be released fro
25. e or similar Gel ar Membrane HDH Gel ar Membrane Gel or Membrane Titerplate or similar Last Modified 13 358 Copy Edit Rename Delete Print Import Export Export All Close Help Figure 14 New Labware Definition 5 Now select HDR Gel 2 x 2 from the list shown in Figure 14 then click the Edit button Edit the parameters of the labware to define a 2 x 2 matrix as shown Figure 15 Biomek 2000 25 HDR User s Guide HDA GEL 2 x 2 e Columns E Grid Spacing Fams 1 25 F r PCLLLLIELLLLL Columns 1 25 mm TT T_T TTT Tet et ee TTT TT TE TET TT TTT TT Well Depth 11 752 mim Tap Height 26 00 Lid Parameters Lid Length Lid Width width B Lid on Height Labware End View Lip Height D Acceleration 1 00 Lid Height IET Below battor Lifter Height F at top Height 0 00 mm Grip Lid Depth 6 Figure 15 New HDR Gel Definition 6 Click OK when done 7 The newly defined gel will appear as an option in the Labware Bar as shown in Figure 16 Function Palette 7 Labware Bar Plates C Tips C Tubes C Devices C Other with Lid Figure 16 Labware Bar With this gel definition each gel will receive four deposits at each grid location 26 Bi
26. e information on using the Aspirate function 5 Replenish the fluid by clicking on the Bulk Dispense icon button on the function palette see Figure 49 then on the desired HDR reservoir Figure 49 Bulk Dispense icon 6 The following window is displayed Figure 50 48 Biomek 2000 HDR User s Guide Document 609930 AA ulk Dispense wass m Figure 50 Dispensing Liquid to a Reservoir 7 Ensure that the correct source is selected if you have a six port valve otherwise the software will assume the single source Enter the parameters for the bulk dispense of liquid using Buffer 2 as your Ethanol source Click OK when done The reservoir will be replenished at the specified point in the method when it is run See the BioWorks User s Manual if you need more information about the Bulk Dispense function Biomek 2000 49 HDR User s Guide Preparing to Run a Replication To prepare to run a replication set up the Biomek Worksurface sterilizer system plates and gels as follows 1 Fill the bleach reservoir with 80 mL of 3 to 12 bleach NOTE The bleach should be drained and properly discarded at the end of each day The tool should be washed with bleach then deionized water after each session Fill the water reservoir with 80 mL of sterile water Fill the ethanol reservoir with 80 mL of 95 ethanol Make sure the drying fan s power supply is connected UT E 2 Place the 96 or 384
27. eight 17 38 E mm Labware End View Lip Height D 2 84 E mm Acceleration 1002 Lid Height E 9 26 mmi Below bottom Lifter Height FI b 33 1 mm Grip at tap Height 0 00 BH mm Grip Lid Depth 61 5 00 mm Figure 9 Specifying the HDR Gel i Enter the number of rows and columns in the matrix This matrix is the same for all grid locations of the Gel As you change the values the matrix graphic changes accordingly The first deposit position at any grid location will always be at the upper left corner Biomek 2000 19 HDR User s Guide Maximum matrix values depend on the sample and the gel media being used However the following maximum matrix sizes are recommended 96 Pin Tool 6x6 0 015 pins or 4x4 0 045 pins 384 Pin Tool 3x3 0 015 pins or 2x2 0 045 pins To change the spacing between the deposit positions of the matrix edit the Grid Spacing values Note the following e When using a 96 pin tool the default spacing must be reduced for matrices of 7 x 7 or greater e When using a 384 Pin Tool make sure that the Matrix value times the Grid Spacing value does not exceed 4 5 mm x 4 5 mm maximum matrix size Overlapping deposits may result The HDR Gel dialog also includes height and depth parameters For Top Height enter the height of the labware measured from the top of the well to the surface of the Biomek For Well
28. en you are finished the method window should look like the one shown in Figure 21 lt BOOK1 gt Method 1 IDEE Initial Configuration Heplicate 44 using HDHT Heplicate 45 to B4 using HDHT Sterilize Replicate using HORT Sterilize Figure 21 Method Screen for Example Method 1 30 Biomek 2000 HDR User s Guide Document 609930 AA Example Method 2 HDR Transfers within Loops In this method four replicate gels are made from four sample plates For this example the operator will be prompted to change the sample plate after replications are made to each of the destination gels 1 Double click on the Edit icon in the BioWorks program group Then select Method New Click Clear Configuration 2 Set up the Initial Configuration of the worksurface as shown in Figure 22 Use the 2 x 2 Gel defined earlier in this section as the destination media We will use a Manual Move command to place the 96 well flat plate for the source Biomek 2000 Worksurface m ngoaggogoagogdgaga LI m
29. he same loop insert another loop to perform an HDR transfer to each of the gels Because there are 16 samples set up the loop for a count of 16 Figure 38 Begin Loop Loop for time 10 00 00 Cancel Loop until operator cancels Help Loop until all labware used Figure 38 Specifying a Loop for Sample Placement 40 Biomek 2000 HDR User s Guide Document 609930 AA 15 Click on Next Labware icon On the next Labware screen select Remove current item get next item Click OK when done 16 Set up an HDR transfer from the sample at B6 to each of the gels configuring a sterilization after the last transfer Figure 39 High Density Replication HDR Transfer Destination Labware Location E Label HEF GEL 4 4 GEL 4 x 4 Pause 1 E sec Source Labware Location Label f6 well flat Type S6 well Pause 1 Rate Tool HORT bi Bleach Water Direction m Ethanal HELE Fan Pause A d v Sterilize Cancel Help Directa Figure 39 HDR Transfer for Example Method 3 17 Insert an End Loop command after the last replication step Next click on the Next Labware icon then click on Worksurface location B6 On the Next Labware screen select Remove Current item Only Then insert another End Loop command 18 The method should now look like the
30. ion Figure 44 Warning Dialog for HDR Fan extending beyond Tool Rack Likewise when you place labware next to the HDRT tool rack position A4 in the example shown you may also receive warnings about pipetting to that position To replenish liquid in a reservoir do the following 1 Purge the line of the Wash tool by clicking on the Purge icon on the Labware Bar then on the wash tool 2 A window similar to the one in Figure 46 is displayed Figure 45 Purge icon 46 Biomek 2000 HDR User s Guide Document 609930 AA Tool Source Reagent Bufferl Buffer Buffers Buffer C Bufferh Purge Volume 10 0002 ml per channel Purge Hate 10 E Cancel Help Figure 46 Purging the Wash Tool Select the source from which you wish to purge the tool Buffer 1 the volume you wish to purge and the rate the liquid is forced through the lines 1 is the slowest 3 Specify an aspirate function by clicking on the Aspirate icon button on the function palette see Figure 47 then click on the reservoir to replenish 4 The following window Figure 48 is displayed Figure 47 Aspirate icon Biomek 2000 47 HDR User s Guide 0002 m Aspat sides Figure 48 Aspirating Liquid from a Reservoir Enter the aspiration parameters and click on OK See the BioWorks User s Manual if you need mor
31. l Fat Titerplate or similar Jb well vinyl u bat Titerplate or similar vinyl Beckman Coming Costar 24 Cryo v bat Falcon 24 Filter Plate FlashPlate Full Reservoir HOA Bleach Ethanol Gel 384 Gel water Microtuge 24 Titerplate similar Titerplate or similar Titerplate similar Titerplate similar Titerplate or similar Tube Module mall Titerplate or similar Titerplate similar Titerplate or similar Titerplate or similar Titerplate similar Titerplate or similar Gel ar Membrane Gel or Membrane Titerplate similar Tube Module mall Rename ELO Bee E Enen EN Delete Frint Import E port Export All lose 00 Titerplate similar Help Figure 8 Selecting the HDR Gel 18 Biomek 2000 HDR User s Guide Document 609930 AA Select HDR Gel or HDR Gel 384 from the list then click the Edit button to display the HDR Gel window Figure 9 ha atris Column Grid Spacing P owa 1 25 E mm 11171 Columna 1 25 mm TOLLLLLLLL LE TE TT TE TTT TT i Well Depth 11 75 mm Tap Height 26 00 2d mm Have Lid Parameters Lid Length 127 11 BH mm Lid width 9910 24 mm Top Width B 31 095 mm Lid on H
32. les use gels with a 2 x 2 deposit matrix for the destination To create this labware copy the default Gel labware and modify it as follows Creating a Custom Gel 1 From the BioWorks editor select the Edit Labware option from the menu bar This displays the Edit Labware dialog shown in Figure 12 E dit Labware Authar keyword Filter Beckman Instruments Gel Library Labware Last Modified Modular Reservoirs pe 9 98 Labware Mame Labware Gel Gel or Membrane Copy J6 well vinyl u bat Titerplate similar Sb well vinyl v bat Titerplate similar a Beckman Titerplate similar Rename Breakaway 9b Titerplate or similar Coming 12 Titerplate similar Delete Costar 24 Titerplate similar Cryo v bat Tube Module small Falcon 24 Titerplate or similar Print Filter Plate Titerplate or similar FlashPlate Titerplate or similar Import Full Reservoir Titerplate similar HD RH Bleach Titerplate or similar E port HOR Titerplate or similar Gel 384 Gel or Membrane HOR HOR Gel or Membrane _ Export al water Titerplate similar Close Microfuge 24 Tube Module small Titerplate similar 4 wall Titerplate or similar 7 Help Figure 12 The Edit Labware Window Biomek 2000 23 HDR User s
33. ll SE wellflat J6 well Figure 30 Populating Stacks with Plates 4 To add gel plates select Plates then go to Labware Bar and select HDR 4x4 Click on Shelves 1 through 5 in Stacks C through F 36 Biomek 2000 HDR User s Guide Document 609930 AA Iu C dn i Erra l I Xu E E INTE IE Ti 1 1 E nmm Figure 31 Populating Stacks with Gels 5 Associate each of the gel positions on the worksurface A4 A5 B4 and B5 with one of the stacks in the stack areas To do this go to the Labware Bar and select the Set Shelf icon as shown in Figure 32 Figure 32 Set Shelf icon 6 Click the Shelf lof Stack C then click on the worksurface position A4 7 The Set Shelf dialog will appear as shown in Figure 33 Click OK when done Biomek 2000 37 HDR User s Guide Figure 33 Specifying the Set Shelf Parameters Use Shelf Cancel Help Destination Location i C1 GEL 4 4 8 Repeat steps 4 5 three more times for the gel positions Your method window should look like Figure 34 CECE r Qus p c 1 Palo ale 111 vial Figure 34 Method screen for Example Method 3 0 38 2000 HDR User s Guide Document 609930 AA
34. m all obligations under all warranties either expressed or implied if any product covered hereby is repaired or modified by persons other than its own authorized service personnel unless such repair by others is made with the written consent of Beckman IT IS EXPRESSLY AGREED THAT THE ABOVE WARRANTY SHALL BE IN LIEU OF ALL WARRANTIES OF FITNESS AND OF THE WARRANTY OF MERCHANT ABILITY AND THAT BECKMAN SHALL HAVE NO LIABILITY FOR SPECIAL OR CONSEQUENTIAL DAMAGES OF ANY KIND FROM ANY CAUSE WHATSOEVER ARISING OUT OF THE MANUFACTURE USE SALE HANDLING REPAIR MAINTENANCE OR REPLACEMENT OF ANY OF THE PRODUCTS SOLD UNDER THIS SALES ORDER Representations and warranties made by any person including dealers and representatives of Beckman which are inconsistent or in conflict with the terms of this warranty shall not be binding upon Beckman unless reduced to writing and approved by an expressly authorized officer of Beckman Biomek 2000 59 HDR User s Guide Document 609930 AA Biomek 2000 software 15 warranted on an as 1s basis without fitness for any specific purpose with the exception of the media that is warranted against defects in materials or workmanship for a period of one year Parts replaced during the warranty period are warranted to the end of the instrument warranty The software is copyrighted and may not be altered or given to a third party without written Beckman authorization Items Not Covered by Warra
35. m falling out when the tool is tipped over during plate replacement Retainer 900122 CDR Figure 51 Retainer 2 3 Inserting the Lay the tool on its side on a flat surface and grasp the pin plate and retainer Remove the 2 Phillips mounting screws turn counter clockwise to release the plate and retainer see Figure 52 To change pins the plate see the next section 54 Biomek 2000 HDR User s Guide Document 609930 AA Tool Body Pin Retainer 900123 CDR Mounting Screws Figure 52 Installing the Pin Plate 4 To attach a plate with pins captured by a retainer reverse the steps above Firmly tighten the 2 Phillips screws 5 Remove the retainer before placing the tool in the Biomek tool rack WARNING Damage to the pins may occur if the tool is used with the retainer in place 6 Store the removed pin plate retainer assembly upside down so it rests on the retainer pins pointing up Biomek 2000 55 HDR User s Guide Replacing Pins in Pin Plate 1 Remove the pin plate from the tool then remove the pin retainer if necessary 2 Place the pin plate on an HDR reservoir with the un beveled end of the 2 mounting holes facing up as shown in Figure 53 3 Drop a pin into each hole starting with the 4 corner pins to keep the plate centered on the reservoir 4 Lay a pin retainer on pins and tighten 2 thumbscrews into the mating attachment holes on the edge
36. ns in which the samples are deposited correspond to 96 well or 384 well plates such that the desired samples may be traced back to the sample of origin The HDR system also offers a sterilization setup which allows the HDRT to be sterilized at any point in the assay The target to which samples are transferred is called a Gel Gel is the Beckman term for any labware and or medium used to receive the transfer Each Gel 1s divided into 96 or 384 grid locations depending on the Gel type 96 position grids have locations measuring 9mm x 9mm 384 position grids have locations measuring 4 5mm x 4 5mm BioWorks is used to define a matrix for each grid location in the Gel The matrix defined in BioWorks indicates the number and pattern of deposit positions and the density of those positions For instance a 96 position Gel with a 4 x 4 matrix would define 16 deposit positions for each grid location totalling 1 536 16 x 96 deposits for the Gel NOTE This manual should be used in conjunction with the BioWorks Oper ating System User s Manual for general instructions on creating and running methods 4 Biomek 2000 HDR User s Guide Document 609930 AA HDR System Components HDRT system is available two configurations 1 96 and 384 systems to accomodate different needs and requirements This allows you to individually select the components that you need In most cases you will need one HDR Tool Body one Plate Kit and
37. ntibiotic resistant bacterial colonies growing at random spots May be caused by wet membranes or agar Allow membranes to desiccate on agar for a few minutes before spotting Stainless steel parts show signs of corrosion May be caused by incomplete rinsing by water after bleach treatment or pins left in bleach Extend the water rinse time and replace worn pins as needed If using the bleach reservoir in the tool rack remove the tool from the bleach and rinse thoroughly after each session Skips step in sterilization Pause may be set to 0 for that sterilization step Change the Pause time the HDR Transfer to a non zero value Colonies too close together The transfer area is too small Increase grid spacing and or decrease the num ber of rows and columns in matrix Switch to 0 015 inch pins if not already in use 52 Biomek 2000 HDR User s Guide Document 609930 AA Overlapping matrices The transfer area is too large Decrease grid spacing and or decrease the num ber of rows and columns in matrix Biomek stalls when picking up the HDR tool Pin retainer plate not removed Stop the method and remove retainer plate General Maintenance Procedures WARNING TURN THE POWER SWITCH OFF AND DISCONNECT THE BIOMEK 2000 FROM THE MAIN POWER SOURCE before attempting any cleaning or maintenance of any component of the Biomek 2000 Introduction The HDR System has been designed to keep customer maintenance sim
38. nty The customer will be charged at standard hourly rates 1f at any time Beckman service is called to perform routine tasks covered in the operators manual such as programming move or reinstall the instrument perform procedures which are covered in the Maintenance section of this manual e work on or diagnose a system problem where the problem is shown to be in a portion of the system not purchased from Beckman repair instrument damage resulting from operator abuse of the instrument work on the instrument but is unable to do so because it requires decontamination which is the operator s safety officer s responsibility train an operator subsequent to initial training during installation Biomek 2000 60 HDR User s Guide Document 609930 AA Glossary of Terms Bleach Reservoir The HDR reservoir identified in the software as the reservoir containing bleach used in the sterilization process The Bleach reservoir may reside in a tool rack or in one of the worksurface locations Fan The fan dries the pins of the HDRT at the end of the sterilization process It must be placed in a tool rack and identified in the BioWorks software Ethanol Reservoir The HDRT reservoir identified in the software as the reservoir containing ethanol used in the sterilization process The Ethanol reservoir may reside in one of the worksurface locations HDR Gel The gel or membrane that receive the sample The structu
39. nual Biomek and Beckman are registered trademarks of Beckman Instruments Inc Microsoft is a registered trademark of Microsoft Corporation Windows is a trademark of Microsoft Corporation Biomek 2000 1 HDR User s Guide Safety Notice The international symbol displayed above and on the equipment is a reminder to the user that all safety instructions should be read and understood before installation operation maintenance or repairs to these instruments are attempted When seeing the symbol on other pages of this manual special attention must be given to the specific safety information which applies If the product is used a manner other than specified in the manual the safety and performance of the equipment could be impaired Summarized safety information basic to the safe operation of the High Density Replicating System HDR on the Biomek 2000 Laboratory Automation Workstation is described below More complete safety information appears in the manual where appropriate e Perform only the maintenance described in the Biomek 2000 Installation and Maintenance Guide and in this manual pertaining specifically to the HDR System Maintenance or repair procedures not specifically described in the above mentioned manuals expose persons to risks of electric shock or injury Refer such servicing to Beckman qualified service personnel The HDR System 16 not autoclavable nor should you use organic solvents to
40. omek 2000 HDR User s Guide Document 609930 AA Example Method 1 Basic HDR Transfer The first example will deposit sample from four individual 96 well plates onto two gels This method will result in two duplicate gels each of which contains samples from all four of the sample plates Labware sample plates and gels are placed on the Biomek worksurface manually Figure 17 shows the pattern used for deposition of the sample on the gel for this example ric ric ric m MOO OOo Figure 17 Sample Deposition on Multiple Gels 1 Open anew method in the BioWorks editor and configure the worksurface as shown in Figure 18 Be sure that you use the 2 x 2 Gel defined in the previous section for the destination labware Biomek 2000 27 HDR User s Guide Biomek 2000 Worksurface b Port Valve Figure 18 Initial Configuration for Sample Method 1 2 To define the HDR Transfer for each of the source plates click on the HDR Tool icon from the Function Palette 2 3 Then click on the first 96 well plate at position 5 then click on the first gel at position A4 The HDR Transfer dialog shown below in Figure 19 is displayed Click OK to accept the parameters 28 Biomek 2000 HDR User s Guide Document 609930 AA
41. p for the Biomek workstation the total system leakage current must not exceed 5 milliamperes values outside of North America may differ Turn the POWER off and disconnect the workstation from the main power source before changing fuses or performing any maintenance NOTICE It is your responsibility to decontaminate components of the Biomek system before requesting service by our Field Service Representative or returning parts to Beckman for repair Beckman will NOT accept any items which have not been decontaminated where it is appropriate to do so If parts are returned they must be clearly labeled with a note stating that they are safe to handle and not contaminated Do not replenish the bleach using the aspirate and bulk dispense transfers as the bleach may damage the Wash tool Do not attempt to use tools other than HDR Tool HDRT when performing a High Density Replicating Transfer Biomek 2000 3 HDR User s Guide Introduction General Description The Biomek 2000 High Density Replicating System HDR System used on the Biomek 2000 Laboratory Automation Workstation is designed for applications which require many small samples to be deposited on a suitable medium for the purposes of growth or analysis The HDR system not only automates this time consuming and error prone process but also allows for the placement of 3 456 or more samples per 8 x 12 cm target area using the HDR Tool HDRT The patter
42. ple The following procedures should be performed routinely Maintenance not covered in this manual should be handled by your Beckman Field Service Representative or other Beckman trained personnel Maintenance contrary to that permitted in this manual or under terms of the warranty may result in damage to the system and invalidation of the warranty agreement see the Warranty HDR System For proper maintenance of the HDR System perform the following on a daily basis sure that you clean the pins and the pin plate section of the tool at the end of each day Use bleach then deionized water to clean these parts This will sterilize the tool The deionized water cleaning is necessary to neutralize the bleach which will cause the stainless steel to corrode if left in contact with bleach during storage e Be sure to empty the bleach and ethanol reservoirs at the end of each day and rinse them out thoroughly with water Biomek 2000 53 HDR User s Guide Replacing the Pin Plate in the HDR Tool Before replacing the pin plate be sure that you have a No 1 small head Phillips screwdriver To remove or replace the pin plate follow the steps below 1 To remove the pin plate hold the tool in its normal operating position and slide the pin retainer into the space just above the tops of the pins Tighten the two captive thumbscrews into the mating holes on the edge of the pin plate see Figure 51 This retainer keeps the pins fro
43. r 384 well plate as the first sample transfer onto the membrane Mix a known amount of control probe e g 1 10 with the specified probe for the hybridization reaction The resulting autoradiogram will have a positive spot in the upper left hand corner of each array Each matrix represents the result of the transfers which are specified in number of rows and columns for each matrix in the software Using this method you can see clearly each matrix so you can locate additional positives Biomek 2000 21 HDR User s Guide Troubleshooting amp Maintenance This section provides a list of possible problems and suggested remedies NOTE General Troubleshooting and Maintenance procedures are described in the BioWorks Installation and Operations Guide Missed transfers on agar or membrane May be caused by liquid dirt or grease residue on pins causing them to stick Clean the pins or pin plate with ethanol and dry thoroughly Check for bent pins Spot size varies larger than normal overnight growth May be caused by grease dirt or other residue on pins or pits on the surface due to corrosion Clean pins with ethanol or replace pins No growth of colonies or inconsistent growth of colonies May be caused by agar which is too dry too low not level or contains air bub bles not enough sample in source plate or ethanol contamination of sample Pour agar so that it is level add more sample to wells increase fan pause times A
44. re of the Gel determines how the transfers can be arranged The Gel is divided into 96 matrices Each matrix receives the same row and column array specified in the BioWorks software HDRT High Density Replicating Tool The tool which transfers 96 or 384 samples at a time from a source plate to a Gel Matrix One of the 96 9 x 9 mm areas in the colony grid which receives the row and column array defined in the BioWorks software Pin Plate The plate that holds the 96 or 384 pins of the HDRT Pins The 96 or 384 transfer pins residing in the pin plate of the HDRT Sterilizer System The Bleach Water Ethanol reservoirs and the fan unit used to sterilize the HDRT Tool Holder This term is also used to refer to the Tool Rack which is a Biomek labware piece used to hold tools on the Biomek worksurface Top Plate The component in the HDRT which pushes the pins down to their original positions before and after each transfer Water Reservoir The HDRT reservoir identified in the software as the reservoir containing water used in the sterilization process The Water reservoir may reside in one of the worksurface locations Biomek 2000 61
45. reservoir is used place the HDRT in the center slot If you are also installing another tool in the rack and the rack is on the left end of the worksurface with no disposal next to it you can use Slot 1 or 2 for the HDRT Be sure to remove the bleach tray if using other tools in the same rack 14 Biomek 2000 HDR User s Guide Document 609930 AA 4 Install the fan unit in one of the tool racks with the fan switch on the left and the two fan tabs sitting in the rack slots as shown in Figure 7 The fan 1s generally installed in Slot 3 If you are also installing other tool s in the rack and the rack is on the left end of the work surface with no disposal next to it the fan may be placed in Slot 1 or 2 of the rack Plug the power supply into the fan unit then route the fan cable using the cable clips if necessary Plug the other end into your power source 900121 CDR Figure 6 Installing the HDRT Biomek 2000 15 HDR User s Guide 900124 CDR Figure 7 The Fan Unit on the Biomek 2000 Worksurface 16 Biomek 2000 HDR User s Guide Document 609930 AA Activating the HDR System in BioWorks To activate the buttons and menu items for the HDR system you must install BioWorks as described in the BioWorks Quick Start Guide and enable the HDR System in the Biomek System Config dialog If you are installing BioWorks for the first time or installing a software upgrade you will indicate that you ha
46. t the minimum requirements specified in the BioWorks Software Reference Manual Migration Guide or Release Notes for the version of BioWorks in use A printer 1s optional however if one is used it should be supported by Windows NT Equipment not recommended by Beckman cannot be installed and or maintained by our Field Service Engineers Optional accessories to the computer such as a printer should be installed according to manufacturer s instructions Facility Requirements Following are the facility requirements for the HDR System accessory Electrical A power supply 110 120V 60 Hz or 220 240V 50 Hz is supplied in each Fan Kit for the operation of the fan used to dry the pins of the HDRT during the sterilization process You must have a single outlet for the power supply in addition to the power supply used for the Biomek system The appropriate outlet should be within 6 ft 2 m of the instrument CAUTION DO NOT use any power supply other than the type supplied for the HDR System as appropriate for the country where the Biomek is installed Space The Biomek 2000 system with the HDR System must be installed in a clean safe and uncluttered environment observing 2 in 50 mm clearances on all sides If using hazardous materials the workstation must be placed in a hood 6 Biomek 2000 HDR User s Guide Document 609930 AA Additional Requirements The necessary supplies for the sterilizer components are blea
47. use Figure 3 4 Placethe tool in the tool rack Do not store the tool with the pin points resting on a flat surface Biomek 2000 11 HDR User s Guide 900122 CDR Figure 3 Removing the Retainer Installing the HDR Sterilizer System Before you can use the HDR System you must install the applicable bleach water and ethanol reservoirs the fan and the HDRT The power supply for the fan is provided Use the steps below to set up the HDR system 1 Install the labware holders and tool racks for the HDRT fan unit reservoirs and labware as described in BioWorks Installation and Maintenance Guide 2 Install the PVC bleach water and ethanol reservoirs in the desired labware racks as shown in Figure 5 12 Biomek 2000 HDR User s Guide Document 609930 AA 900119 CDR Figure 4 Installing Reservoirs into Labware Holders If you install the Bleach reservoir into the Tool Rack use the reser voir supports to hold the reservoir at the right level for the HDRT as shown in Figure 5 You cannot install other tools in the same tool rack if a bleach reservoir is installed Biomek 2000 13 HDR User s Guide 900120 CDR Figure 5 Installing the Bleach Reservoir into the Tool Rack 1 Reservoir Support 3 Tool Rack 3 Install the HDRT into the tool rack with the arrow pointing towards you as shown in Figure 6 If the HDRT 15 the only tool in the rack or a bleach
48. ve an HDR System when specifying the system components during the installation process If you already have BioWorks installed click on the System Config icon in the BioWorks Program Group then enable the High Density Replicating System in the Biomek Setup window The HDR system items will be activated Using the BioWorks HDR Options Introduction The HDR System in BioWorks allows you to create and run customized methods to transfer samples directly onto Gels using the HDRT Gel is the term for any labware and or medium used to receive the transfer The Gel may be a gel or a membrane Gels are divided into 96 or 384 grid locations Each grid location contains an identical definable matrix of deposit positions where the HDRT transfers samples An HDR Transfer Function specified in a BioWorks method involves one or more transfers by the HDRT from the source to the Gel Each transfer makes one deposit at each of the grid locations one pin per grid location per transfer With each subsequent transfer the deposit positions are shifted until the matrix 2 x 2 4 x 4 etc is fulfilled For example a 96 pin transfer for a Gel with a matrix of nine deposit positions at each grid location 3 x 3 matrix requires nine transfers to fulfill the matrix This will generate a Gel with 9 x 96 864 total deposits The same transfer with a 384 Pin tool will make 3 456 deposits To control the number of deposits within a grid location and the spa
49. well plate s containing the samples on the Biomek Remove the lid s if any 6 Place the Gel s on the Biomek Each gel can be either a gel or a gel with membrane Remove the lids if any from the gel labware If using membrane on the target gels the following steps are suggested a Prewet the membrane s in double distilled sterile water b Soak the membrane s 5 to 10 minutes in the same liquid media minus agar c Once the membrane s have been placed on the gel s allow to desiccate for about 5 minutes WARNING If using a Wash tool to replenish the water and or ethanol place the Wash tool reservoir snugly onto the support block If it is not placed properly onto the block the Wash tool may be damaged when the Biomek uses the tool See the Biomek 2000 Installation and Operations Guide for more information Running an HDR Method To run an HDR method double click on the Run icon in the BioWorks Program Group Click on Method Open A list of methods is displayed Double click on the desired method You must verify the physical locations of the HDR setup then the method will run 50 Biomek 2000 HDR User s Guide Document 609930 AA Locating Specific Clones In order to locate positives on the autoradiogram following hybridization we recommend the following methods 1 Cut the membrane to fit the Gel exactly We suggest you cut the bottom right hand corner of the membrane 2 Use a positive control 96 o
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