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2200 TapeStation - User Manual

1. 1000 bp 10 Sensitivity 0 05 ng pL Sizing Precision 5 CV Sizing Accuracy 10 Quantitative Precision 10 CV Quantitative Accuracy 20 Quantitative Range 0 1 50 ng L Carry Over N A Physical Specification Analysis Time Samples per consumable Sample Volume Required Shelf Life Box Kit size 16 samples lt 20 min 96 samples lt 100 min 16 Tul 4 months 112 samples box 1 2 Signal noise ratio gt 3 for single peak 3 Determined using the D1K ladder as sample Agilent 2200 TapeStation User Manual Resolution is defined as the separation of fragments at half peak height or better 15 2 16 Specifications Specification Genomic DNA ScreenTape Assay Analytical Specification Genomic DNA ScreenTape Sizing Range Sensitivity Sizing Precision Sizing Accuracy Quantitative Precision Quantitative Accuracy Linear Concentration Range Carry Over Physical Specification Analysis Time Samples per consumable Sample Volume Required Shelf Life Box Kit size 200 bp to gt 60000 bp 0 5 ng pL 200 15000 bp 15 CV 200 15000 bp 10 15 CV 20 10 100 ng L N A 16 samples lt 25 min 96 samples lt 150 min 16 Tul 4 months 112 samples box 1 Determined using the Genomic DNA ladder as sample 2 Average result from various genomic DNA sample types Agilent 2200 TapeStation User Manual Specifications 2 Specification High Sensitivity R6K S
2. The resultant stain solution should be thoroughly mixed before use For certain applications particularly with high protein concentrations higher concentrations of stain can be used in combination with altered ratios of stain to sample a 16 samples prepare 40 uL stain solution v 2 Stain protein sample or ladder The P200 ladder should be processed through the P200 sample preparation procedure in the same manner as your samples In Ladder mode selected in the ladder options in the controller software P200 ladder is automatically selected as the first sample in the TapeStation controller The user can also select to run no ladder and then to insert a software saved ladder in the 2200 TapeStation Analysis software Agilent 2200 TapeStation User Manual 63 4 64 Using the 2200 TapeStation System a Place 2 uL of P200 stain solution prepared above into a PCR tube strip or 96 well plate b Pipette 2 uL of the protein sample or ladder into the tube mix and attach the lids or foil cover to prevent evaporation c Heat for 7 min at 75 C d After heating remove condensation from the lids or foil cover of the tubes by centrifugation P200 pH buffer clear is supplied to allow the user to dilute samples to alleviate issues with staining efficiency caused by low pH The use of P200 pH Buffer resolves these issues in most circumstances For further information on buffer compatibility contact your Agilent
3. Prepare samples according to type as detailed in How to prepare your samples on page 47 or the appropriate Quick Guide 8 Place samples into the sample block inside the TapeStation Damage to the 2200 TapeStation and impact on performance gt Ensure the lids have been removed from the sample tubes 38 9 Select the tubes or wells you wish to run by clicking and dragging the mouse over the sample locations Selected wells will change colour from white Selected lanes on the controller ScreenTape image will change colour e Lanes which have been run previously will appear grey For best sizing precision and accuracy the user should run the appropriate ladder with the samples For RNA applications or if 16 samples need to be analysed in parallel the user may insert a saved ladder in the 2200 TapeStation analysis software RNA reagent kits do not contain a ladder No software saved ladder is available for genomic DNA applications ScreenTape can be used up to 2 weeks after first use if it has been stored upright between 2 8 C Simply select the samples in the same manner as whole ScreenTape The first sample selected will automatically appear in the first available lane Partially used ScreenTape those that contain lanes run on previous occasions should be returned to the box and stored vertically between 2 8 C for a maximum of 2 weeks D1K Genomic DNA and R6K Reagents Store between 2 8 C P200 R
4. Technologies representative 3 Denaturate sample a Choose which sample buffer is required P200 Reducing Sample Buffer O or P200 Non reducing Sample Buffer It is recommended that P200 Reducing Sample Buffer is used for the denaturation of P200 Ladder b Add 4 uL of the relevant P200 sample buffer to the stained sample and replace the lids or foil cover c Mix and heat at 75 C for 5 min d Remove condensation from the lids or foil cover of the tubes by centrifugation 4 Add 2 uL of P200 Marker to each sample and to the P200 ladder 5 Mix the solution well and centrifuge to ensure that the sample is at the bottom of the tube ready for analysis on the TapeStation P200 Marker is formulated with a high percentage of glycerol Due to the high density of this reagent the user must ensure that the samples are adequately mixed prior to analysis on the TapeStation Failure to do so may result in unsatisfactory analysis results Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Protein Sample Preparation 6 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section gt gt saie or P200 Ladder Vg PL Viz 2 pL V 8pL tot Agilent 2200 TapeStation User Manual 65 4 Using the 2200 TapeStation System Protein Sample Preparation 66 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation Us
5. be copied and pasted from for example an Excel table The entered Sample descriptions data can be copied to clipboard by using the Copy to Clipboard link in the top right hand corner of the Sample Description table Agilent 2200 TapeStation User Manual 41 4 Using the 2200 TapeStation System Tools Protocol Ladder Needle Change Transport Help TapeStation User Agilent Technologies Sample Descriptions 0100100101010 010101010101010 D1K ScreenTape D gt Agilent Technologies ID 00 S019 010102 99 121287 Expires 28 May 2040 BP ik Seranta Figure 5 Controller image 8 way strip selection Protocol Ladder Needle Change Transport Help Agilent Technologies 10 0 elelejelelelelejele 2 Sampie Well Description 300 O E a u In the 96 well plate sample selection screen the panel labeled LDR is no longer available for selection All information entered in the control software will appear in the analysed results 42 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Start the TapeStation Run 1 Click the start button This will produce a Save As window As a default the file name starts with the date in reverse order and a run counter When run continuously the save function auto increments the counter part of the file name yore Ep 2200 TapaSusion Dats File pone ott 1014 09 Save or pe PAF T apeStaton O ata fe l AEF 2 Type in the name that you wish the a
6. e 2200 TapeStation Technical Specification e 2200 TapeStation Components Agilent 2200 TapeStation User Manual Introduction to the 2200 TapeStation System 1 e Installing the 2200 TapeStation Information pertaining to sample and ScreenTape requirements can be found in e ScreenTape Architecture e Operating Procedure ScreenTape Architecture Barcode Buffer chamber Electrodes Gel ScreenTape product details Front View s ScreenTape details 4 r 4 i 4 a n 1 4 r 4 45 r 15 a 5 h 4 s ot H Buffer Chamber gt 1 5 1 5 a 1 7 ea H a a Ss a a t lt Electrodes gt 4 1 r at 4 r a 1 1 Gel gt gt a t 4 Figure 1 Example P200 ScreenTape Agilent 2200 TapeStation User Manual The unique barcode tracks lane usage within individual ScreenTape and allows traceability of results The buffer chamber is located at the top of the channel and contains optimised buffers for the effective separation of nucleic acid fragments or proteins The integrated electrodes apply a current across the ScreenTape and eliminate the need for any additional electrophoresis equipment The gel contained within ScreenTape has been developed specifically to resolve nucleic acids or proteins The information is unique to each consumable item This includes ScreenTape type product expiry date and a unique serial number Back View iD 00 58
7. solvents samples and reagents can hold health and safety risks gt When using handling the ScreenTape and working with these substances observe appropriate safety procedures for example by wearing goggles safety gloves and protective clothing gt Always follow good laboratory practices and adhere to the guidelines established in your laboratory gt Refer to product material safety datasheets for further information gt The volume of substances should be reduced to the minimum required for the analysis CAUTION Damage to the 2200 TapeStation instrument gt Use only the recommended consumables and reagents with the 2200 TapeStation system When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to viscosity of Sample Buffers e When pipetting small volumes ensure that no sample remains within the tip When adding sample buffer to sample please ensure that they are mixed correctly To achieve this gently mix several times with additional pipetting then cap the tubes mix the samples using a vortex mixer for 5 s and briefly centrifuge to collect the contents at the base of the tubes Improper mixing can lead to quantification errors For best results ensure that all reagents are allowed to equilibrate to room temperature for 30 minutes prior to use Agilent 2200 TapeStation User Manual 47 4 48 Using the 2200
8. the instrument at your site and initiate appropriate actions Personal injury The TapeStation is heavy gt Enlist the aid of a co worker to share the lifting load to avoid personal injury 22 1 Remove the TapeStation from the packaging and place on a clean dry flat surface 2 Allow the TapeStation to acclimatise to the ambient temperature of the operating environment Agilent 2200 TapeStation User Manual Installing the System 3 3 Remove the label covering the tape holder as shown in the image below Figure 4 Remove before use Delivery Checklist Ensure all parts and materials have been delivered with your system The delivery checklist is shown below Please report any missing or damaged parts to your local Agilent Technologies sales and service office Agilent 2200 TapeStation User Manual 23 3 Installing the System Contents of the ScreenTape System The Agilent 2200 TapeStation Table1 The Agilent 2200 TapeStation System G2964AA G2965AA Product Volume Properties Agilent 2200 1x Instrument for loading electrophoresing imaging and analysing TapeStation 2200 TapeStation System G2964AA or e 2200 TapeStation Nucleic Acid System G2965AA TapeStation Software 1xCD The software is required to drive the 2200 TapeStation and visualise the Setup Disc ScreenTape analysis Laptop 1 x Laptop Instrument Control Laptop USB Cables Power 1 x USB cable 1x USB cable to connect the laptop to the Tape
9. 0 4 1 Introduction to the 2200 TapeStation System Agilent 2200 TapeStation Components Lid The 2200 TapeStation lid must be closed each time the instrument controller software is initialized and whilst the instrument is in operation LED The LED will illuminate once the instrument is on When the LED is flashing slowly the instrument is in use and the lid should not be opened rapid flashing indicates that the TapeStation requires some attention Sample Block There are 2 sample blocks provided that can either hold 0 2 mL sample tube strips or a 96 well plate Tip Holder The tip holder can accommodate up to 16 TapeStation loading tips at any one time ScreenTape The tape must be placed into the holder with the barcode towards the front of the instrument facing towards the right USB Socket The USB connector is inserted into the USB socket to link the laptop to the 2200 TapeStation Power cable socket The power cable must be connected to the 2200 TapeStation and the relevant mains electricity outlet 8 Agilent 2200 TapeStation User Manual Introduction to the 2200 TapeStation System 1 Lid ScreenTape Tip holder Sample block LED Figure 2 2200 TapeStation front view On off switch Power cable socket USB socket Figure 3 2200 TapeStation back view Agilent 2200 TapeStation User Manual 1 Introduction to the 2200 TapeStation System Overview of the System 10 Agilent 220
10. 0 TapeStation User Manual a Agilent 2200 TapeStation User Manual KP amp e o o e00 e 72 7 e Specifications e e e Technical Specifications 12 ScreenTape Specifications 14 This chapter provides information on specifications i Agilent Technologies 2 Specifications Technical Specifications 2200 TapeStation Input voltage Power consumption Current Interface Instrument Housing Dimensions Weight Power Supply Input voltage Input frequency Phase Current 12 VDC 40 W VA 3A USB cable PC comms UL94 V0 rated flame retardant cast polyurethane 400 x 310 x 310 mm 12 5 kg 100 240 V AC 50 60 Hz 1 0 45 11A Environmental condition Optimal operating temperature Ambient operating temperature 23 C 73 4 F 12 37 C 53 6 98 6 F for DIK 17 37 C 62 6 98 6 F for HS D1K 15 30 C 59 0 86 0 F for Genomic DNA 14 30 C 57 2 86 0 F for RNA applications 10 33 C 50 0 91 4 F for Protein applications 12 Agilent 2200 TapeStation User Manual Specifications 2 If the instrument is out of the recommended temperature range for the ScreenTape inserted the following error message will appear in the controller software Sample Descriptions Semple Well Oesenpton DIK ScreenTape 10 00 S019 010102 99 121311 If the quoted current temperature is above the specified range pleas
11. 7 ST019 ST010 G2960A G2961A G2964AA G2965AA and G2966AA New needles cartridges can be ordered at any time from Agilent Technologies by contacting your local sales agent For details on correct needle cartridge for your TapeStation model refer to Table 2 on page 69 Change the needle cartridge 1 Remove the sample plate and tip holder 2 Remove the foil tab from the top of the needle cartridge Care must be taken to keep the needle cartridge level after removing the foil tab 3 Insert the needle cartridge into the tip holder space using the label for orientation The cartridge should be placed so that the label faces to the right and the printed arrow points to the front of the TapeStation Rear m Front TapeStation Needle cartridge 4 Close the lid 5 Go to Needle Change on the Controller software toolbar and select Run Agilent 2200 TapeStation User Manual 71 Changing the Needle 72 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual 6 Appendix Limited Use Label License 74 Sound Emission 75 The Waste Electrical and Electronic Equipment Directive 76 Technical Service 77 This chapter provides addition information Apg Agilent Technologies 73 Appendix Limited Use Label License 74 Some products within this system contain SYBR Green I or SYBR Gold nucleic acid stain which are provided under an agreement between Molecular Probes Inc a wholly owned
12. Agilent 2200 TapeStation gn e Ps he 2 amp Qo PR Pd gt i e gt Say 5 BERS Orr f Lh os lt K gow User Manual Agilent Technologies Notices Agilent Technologies Inc 2011 2012 No part of this manual may be reproduced in any form or by any means including elec tronic storage and retrieval or translation into a foreign language without prior agree ment and written consent from Agilent Technologies Inc as governed by United States and international copyright laws Manual Part Number 62964 90001 Edition 11 2012 Printed in Germany Agilent Technologies Hewlett Packard Strasse 8 76337 Waldbronn This product may be used as a com ponent of an in vitro diagnostic sys tem if the system is registered with the appropriate authorities and com plies with the relevant regulations Otherwise it is intended only for gen eral laboratory use Warranty The material contained in this docu ment is provided as is and is sub ject to being changed without notice in future editions Further to the max imum extent permitted by applicable law Agilent disclaims all warranties either express or implied with regard to this manual and any information contained herein including but not limited to the implied warranties of merchantability and fitness for a par ticular purpose Agilent shall not be liable for errors or for incidental or consequential damages in connec
13. Station supply units 2 x power cords 1 x Power supply unit for the laptop 1 x Power supply unit for the TapeStation Sample Block 1 x 0 2 mL strip A removable sample block for the correct loading of samples within the and TapeStation 1 x96 well plate Tip Holder 2x A removable cartridge for pipette tips placed in the TapeStation TapeStation loading 1 x 384 tips Pipette tips to use in the 2200 TapeStation tips TapeStation 1x box of 120 Tube strips for placing samples mixed with loading buffer into the 2200 compatible 0 2 mL tube strips and lids 96 well plates 96 well plate foil seal Loading tip transfer tool optional Guides tubes and caps pack of 10 pack of 100 1x TapeStation Site Safety guide and Quick Guides G2964AA Protein DNA and RNA G2965AA DNA and RNA 24 Agilent 2200 TapeStation User Manual Installing the System 3 ScreenTape Products Kit Components High Sensitivity D1K Assay Part Number Name Color Amount 5067 5363 High Sensitivity D1K ScreenTape 7 ScreenTape 5067 5364 High Sensitivity D1K Reagents 2 vials e High Sensitivity D1K Ladder e 75 pL e High Sensitivity D1K Sample Buffer 300 pL Kit Components D1K Assay Part Number Name Color Amount 5067 5361 D1K ScreenTape 7 ScreenTape 5067 5362 D1K Reagents 2 vials DIK Ladder 75 pL D1K Sample Buffer 400 uL Kit Components Genomic DNA Assay Part Number Name Color Amount 5067 5365 Genomic DNA Screen
14. TAPESTATION INSTRUMENT CONTROL AND DATA ANALYSIS Agilent 2200 TapeStation User Manual 27 3 Installing the System Agilent 2200 TapeStation Set Up Hardware required Software required Laptop Agilent 2200 TapeStation Software already installed Personal injury explosion or fire gt Do not operate the instrument in an atmosphere containing explosive gases or near flammable volatile liquids gt Only approved mains cord set supplied with the instrument must be used with this instrument and if an extension lead is required the lead must be earthed gt lf the equipment is used in a manner not specified by the manufacturer the protection provided by the equipment may be impaired For general safety information please refer to the 2200 TapeStation System Site and Safety Manual Use of unsupplied cables or power adaptors Using cables or power adaptors not supplied by Agilent Technologies can lead to damage of the electronic components or personal injury gt Never use cables or power adaptors other than the ones supplied by Agilent Technologies to ensure proper functionality and compliance with safety or EMC regulations 28 1 Connect the supplied USB cable between the port on the back of the instrument and your laptop 2 Power the instrument with the supplied power lead and adaptor 3 Turn the instrument on using the power switch located at the back of the TapeStation When powered and idle the instru
15. Tape 7 ScreenTape 5067 5366 Genomic DNA Reagents 2 vials Genomic DNA Ladder e 75 pL Genomic DNA Sample Buffer O 1350 pL Agilent 2200 TapeStation User Manual 25 3 Installing the System Kit Components High Sensitivity R6K Assay Part Number Name Color Amount 5067 5369 High Sensitivity R6K ScreenTape 7 ScreenTape 5067 5370 High Sensitivity R6K Reagents 1 vial High Sensitivity R6K Sample Buffer 300 pL Kit Components R6K Assay Part Number Name Color Amount 5067 5367 R6K ScreenTape 7 ScreenTape 5067 5368 R6K Reagents 1 vial R6K Sample Buffer 500 uL Kit Components P200 Assay Part Number Name Color Amount 5067 5371 P200 ScreenTape 7 ScreenTape 5067 5372 P200 Reagents P200 5X Labeling Dye 70 pL P200 Labeling Buffer O 350 uL P200 Reducing Sample Buffer O 550 pL P200 pH Buffer clear 1000 pL P200 Non Reducing Sample Buffer 550 uL P200 Markers pre stained e 270 pL P200 Ladder 40 uL 26 Agilent 2200 TapeStation User Manual Installing the System 3 Installing the System Software Installation The software for your Agilent 2200 TapeStation system is preinstalled on the system laptop For updates or if you have to change the laptop you may download the latest version of the software from the update server http www agilent com genomics tapestation For details on installation of the software refer to the readme txt file on the installation CD Agilent 2200 TapeStation Software
16. TapeStation System For successful loading the sample solution must be placed at the bottom of the tube or well without any air bubbles The 2200 TapeStation will load a sample from a minimum of 3 uL onto ScreenTape Ladder Options In Ladder mode in the controller software a ladder should be loaded into the first available lane Alternatively the user can choose to run a software ladder This is done by choosing No ladder in the 2200 TapeStation Controller software ladder menu then running the instrument as normal A software saved ladder can then be inserted in the 2200 TapeStation Analysis software Ladders not run in the first available position or in No ladder mode can later be assigned as ladder using the 2200 TapeStation Analysis Software Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation Quantification Protocol Ensure that sample and sample buffer volumes are from the correct protocol Ensure that the reagents are used with the corresponding ScreenTape type Ensure correct pipetting technique When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Correct Mixing Sample and sample buffer must be vortex mixed on maximum speed for 5 seconds followed by centrifugation to remove any bubbles Insufficient mixing can cause discrepancies in quantifi
17. all reagents are allowed to equilibrate to room temperature for 30 minutes prior to use When using 96 well plates the use of a 96 well plate vortex adaptor is advised to ensure correct sample mixing Improper mixing can lead to quantification errors As with samples in PCR strips briefly centrifuge after vortexing to collect the contents at the base of the tubes before placing into the TapeStation For successful loading the sample solution must be placed at the bottom of the tube or well without any air bubbles The 2200 TapeStation will load a sample from a minimum of 3 uL onto ScreenTape For best sizing precision and accuracy the user should run the appropriate ladder with the samples If 16 samples need to be analysed in parallel the user may choose to insert a saved ladder in the 2200 TapeStation analysis software Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Protein Sample Preparation Sample Preparation P200 Assay Parts required NOTE p n Description 5067 5372 P200 Reagents 1 Prepare the P200 stain solution a Dilute P200 5X Labeling Dye at a ratio of 1 5 with P200 Labeling Buffer The prepared stain solution is best used on the day of formulation however it can be stored for up to one week below 20 C For normal applications 2 uL of formulated stain solution is required for 2 uL of sample For 16 samples 8 uL of 5X Stain would be diluted with 32 uL of Stain Buffer
18. ample Buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to the viscosity of Sample Buffers e When pipetting small volumes ensure that no sample remains within the tip e Please ensure samples and Sample Buffer are mixed correctly To achieve this gently mix several times with additional pipetting then cap the tubes gently vortex mix for 5 s followed by briefly centrifuging on maximum speed to collect the contents at the base of the tubes This is essential for accurate quantification of samples For best results ensure that all reagents are allowed to equilibrate to room temperature prior to use RNA applications are only available to run without a ladder If required a software ladder can be added in the Agilent 2200 TapeStation analysis software Agilent 2200 TapeStation User Manual 59 4 Using the 2200 TapeStation System Sample Preparation High Sensitivity R6K Assay Description Parts required p n High Sensitivity R6K Reagents 1 5067 5370 1 Mix 1 uL High Sensitivity R6K Sample Buffer with 2 uL RNA sample 2 Sample denaturation Heat the samples to 72 C for 3 min Place samples on ice for 2 min Briefly centrifuge the samples to collect the contents in the base of the tubes 3 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section Ice for 2 min High sensitivit
19. apeStation for Protein Combined TapeStation G2960A 2200 TapeStation System G2961A 2200 TapeStation Nucleic Acid System G2964AA 2200 TapeStation System Twin G2960 60063 G2965AA 2200 TapeStation Nucleic Acid System G2966AA 2200 TapeStation ScreenPlex System Agilent 2200 TapeStation User Manual 5 69 5 Maintenance Needle change intervals e After 3840 7680 lanes in a Dual loading system pierces the controller software will inform the user that a needle change is pending The word Needle will appear in the bottom of the controller software inside a yellow box e After 4160 pierces 8320 lanes in a Dual loading system a needle change is recommended The box around the word Needle will change from yellow to red e After 4480 pierces 8960 lanes in a Dual loading system the needle has completed its lifetime and must be changed before the TapeStation will start leedle Change Transport Help TapeStationUser Agilent Technologies 1 Sample Descriptions Sample Well Description Please insert a ScreenTape Ready Figure 10 Controller software indicating a Needle change is recommended 70 Agilent 2200 TapeStation User Manual Maintenance 5 Parts required p n Description 1 G2960 60062 Needle cartridge for use in single pump systems For use with product numbers ST007 ST008 and ST009 OR 1 G2960 60063 Needle cartridge for use in dual pump systems For use with product numbers ST01
20. cation Mixing Test D1K 120 oC os 100 c Soc 80 aoo Bx Se 60 4 Soe 7 oo oge 40 oto Bo o2 20 gt a m 04 5 Vortex mixing then spin down Pipette mixing only No mixing Figure 7 Effect of sample mixing on quantification results Agilent 2200 TapeStation User Manual 49 4 50 Using the 2200 TapeStation System Peak Integration Ensure that the upper marker is properly integrated This is used for quantification Sample peaks should also be adjusted as required see Figure 8 on page 50 Sample A1 correct Sample A1 incorrect peak integration peak integration Concentration 42 2 ng pL Concentration 29 9 ng pL i S CF mij Figure 8 Peak integration examples for correct and incorrect integration Quantitative Range For accurate quantification ensure that the sample is within the range of the chosen ScreenTape e The quantitative range for D1K ScreenTape is 100 pg uL 50 ng uL e The quantitative range for HSD1K ScreenTape is 75 1000 pg uL In extreme cases overloading the ScreenTape will result in a loss of the bottom marker Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Other Issues Residual AMPure beads from SureSelect protocol can give signal which runs with the upper marker see figure below Any signal under the upper marker affects quantification Removal of the beads removes the signa
21. creenTape Assay Analytical Specification High Sensitivity R6K ScreenTape Assay Quality Score RINE Sensitivity 100 pg pL Quantitative Precision 20 CV Qualitative Range 100 10000 pg pL Physical Specification Analysis Time 16 samples lt 15 min 96 samples 100 min Samples per consumable 16 Sample Volume Required 2 ul Shelf Life 4 months ScreenTape box size 112 samples box 1 Within a ScreenTape Agilent 2200 TapeStation User Manual 17 2 18 Specifications Specification R6K ScreenTape Assay Analytical Specification R6K ScreenTape Assay Quality Score Sensitivity Quantitative Precision Qualitative Range Physical Specification Analysis Time Samples per consumable Sample Volume Required Shelf Life ScreenTape box size RIN 2 ng pL 15 CV 2 500 ng L 16 samples lt 20 min 96 samples 100 min 16 Tul 4 months 112 samples box 1 Within a ScreenTape Agilent 2200 TapeStation User Manual Specifications 2 Specification P200 ScreenTape Assay Analytical Specification P200 ScreenTape Assay Sizing range 10 200 kDa Resolution 15 Typical Sizing Accuracy Sizing Precision Quantitative Range precision Qualitative Range Sensitivity Physical Specification Sample volume needed Analysis Time 10 CAII Lysozyme beta lactoglobulin 3 CV 100 1000 ng pL for IgG 15 CV 5 5000 ng L BSA Lysozyme 12 5 5000 ng L IgG 5
22. e Buffer with 1 uL DNA sample by vortex for 5 s 3 Spin down to position the sample at the bottom of the tube 4 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section cr oat B 1uL sample 1 uL sample 3 uL sample buffer EA 56 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System Sample Preparation Genomic DNA Assay Parts required p n Description 5067 5366 Genomic DNA Reagents 1 Equilibrate all reagents to room temperature for 30 min 2 Prepare Ladder a Aliquot a minimum of 3 uL Genomic DNA Ladder into the first tube well Use a fresh ladder for each run If using 96 well plates always run the ladder in first selected position No software saved ladder is available for the Genomic DNA assay 4 Do not shake or over vortex ladder vial This could result in degradation of the gDNA ladder 3 Prepare Sample a Mix 1 uL DNA sample with 10 uL Genomic DNA Sample Buffer b Spin down then vortex for 5 s c Spin down to position the sample at the bottom of the tube 4 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section Genomic DNA if 10 es D Ti vortex and spin v gDNA sample 1 uL Total volume 11 uL we Agilent 2200 TapeStation User Manual 57 4 Using the 2200 TapeStation System RNA Sample Preparation Information on Working with RNA CAUTION Sam
23. e User Additional Consumables required for the 2200 TapeStation instrument e Loading tips 5067 5152 or 5067 5153 Optical Tube 8x Strip 401428 and and Optical Cap 8x Strip 401425 or 96 well Sample Plates 5067 5150 and 96 well Plate Foil Seal 5067 5154 Additional Material Required Not Supplied e Volumetric pipette e Vortex mixer Centrifuge Heating block or PCR machine 34 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Operating Procedure 1 Double click the 2200 TapeStation controller icon on the desktop and follow the instructions on the screen Always ensure you are using the most up to date Controller software Please check for the latest version You will now see the startup splash TapeStation Software Controller 2 Insert the tube strip sample block into the TapeStation gt 7 p OR Insert the plate sample block into the TapeStation 2P Agilent 2200 TapeStation User Manual 35 Using the 2200 TapeStation System Operating Procedure 3 Place loading tips into the loading tip holder as shown and insert into the TapeStation EN be removed The 2200 TapeStation will not run until all the tip buckets are empty NOTE If any used tips are left in the tip buckets a pop up window will ask for the discarded tips to NOTE A Loading tip transfer tool G2964 60000 is available NOTE Ensure that all 16 loading tips are inse
24. e move the system out of direct sunlight and away from any windows Check that any air conditioning is functioning If the quoted current temperature is below the specified range please allow the instrument to equilibrate to the ambient temperature and avoid using in a cooled area Agilent 2200 TapeStation User Manual 13 2 ScreenTape Specifications 14 Specifications Specification High Sensitivity D1K ScreenTape Assay Analytical Specification High Sensitivity D1K ScreenTape Assay Sizing Range Resolution Sensitivity Sizing Precision Sizing Accuracy Quantitative Precision Quantitative Accuracy Quantitative Range Carry Over Physical Specification Analysis Time Samples per consumable Sample Volume Required Shelf Life Box Kit size 35 1000 bp 35 300 bp 15 300 1000 bp 10 5 pg pL 5 CV 10 15 CV 20 75 1000 pg pL N A 16 samples lt 20 min 96 samples lt 100 min 16 2 uL 4 months 112 samples box 1 2 Signal noise ratio gt 3 for single peak Resolution is defined as the separation of fragments at half peak height or better 3 2200 TapeStation Nucleic Acid System G2965AA Determined using the D1K ladder as sample Agilent 2200 TapeStation User Manual Specifications 2 Specification D1K ScreenTape Assay Analytical Specification D1K ScreenTape Assay Sizing Range 35 1000 bp Resolution 35 300 bp 15 300
25. eagents Store from 30 to 20 C Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 10 The sample selection can be deleted by right clicking on the sample plate image A menu will appear with the following options Clear All Selections this will clear ladder well and all sample wells selected e Clear Last Selection this will only clear the last samples to be highlighted Pressing Escape on the keyboard will also cancel the current selections Agilent 2200 TapeStation User Manual 39 4 Using the 2200 TapeStation System Add Experiment Notes 1 If required notes can be manually entered into the software before the instrument is started Tools Protocol Ladder NeedleChange Transport Help TapeStation User Ef Agilent Technologies Sample Descriptions Sample Well Description O Oo O Oo 0O O 0 Oo oO Oo Oo B c D E F G H D1K ScreenTape Agilent Technologies ID 00 S019 010102 99 121287 Expires 28 May 2040 40 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Describe Samples 1 Sample descriptions can be manually entered into the software before the instrument is started and whilst the TapeStation is operating before analysis software is launched OR If samples are barcoded place the cursor in the desired track description and scan the sample barcode with a hand held barcode scanner OR Sample data can
26. er Manual e e ee Maintenance e e e e General Information 68 4 Changing the Needle 69 This chapter describes the maintenance of the TapeStation system ea Agilent Technologies 67 5 Maintenance General Information 68 Annual Preventative Maintenance PM is essential for the TapeStation as it has many moving parts This PM should be arranged with your local Agilent representative and consists of e Fan Filter replacement e Needle replacement Electrophoresis probe replacement e Internal instrument inspection for wear foreign objects and general clean inside and out In addition to the above the engineer will check that the instrument is functional by running a full ScreenTape For customers with exceptionally high usage the needle replacement procedure as detailed in the section below can also be performed between annual PM services Agilent 2200 TapeStation User Manual Changing the Needle Table2 Overview TapeStation Configuration Needle Cartridge Maintenance It is important to know which TapeStation system you have before changing the needle s in order to purchase the correct needle cartridge Product Number TapeStation Configuration Pump Needle Cartridge Ordering Code ST007 TapeStation for ScreenPlex ST008 TapeStation for DNA Single G2960 60062 ST009 TapeStation for Nucleic acids ST017 TapeStation for ScreenPlex ST019 TapeStation for Nucleic acids Twin G2960 60063 ST010 T
27. h size and quantity NOTE Errors in sizing and quantification will result in erroneous molarity calculations 4 Always ensure that the good measurement practices for sizing and quantification have been followed to ensure accurate molarity values Agilent 2200 TapeStation User Manual 53 4 Using the 2200 TapeStation System DNA Sample Preparation 54 Information on Working with Samples e When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to viscosity of Sample Buffers e When pipetting small volumes ensure that no sample remains within the tip e When adding sample buffer to sample please ensure that they are mixed correctly To achieve this mix several times with additional pipetting then cap the tubes mix the samples using a vortex mixer on maximum speed for 5 s and briefly centrifuge to collect the contents at the base of the tubes Improper mixing can lead to quantification errors e For best results ensure that all reagents are allowed to equilibrate to room temperature for 30 minutes prior to use When using 96 well plates the use of a 96 well plate vortex adaptor is advised to ensure correct sample mixing Improper mixing can lead to quantification errors As with samples in PCR strips briefly centrifuge after vortexing to collect the contents at the base of the tubes before placing into the TapeStation F
28. his product for purposes other than research contact Molecular Probes Inc Business Development 29851 Willow Creek Road Eugene OR 97402 USA Tel 541 465 8300 Fax 541 335 0354 Agilent 2200 TapeStation User Manual Appendix 6 Sound Emission Manufacturer s Declaration This statement is provided to comply with the requirements of the German Sound Emission Directive of 18 January 1991 This product has a sound pressure emission at the operator position lt 70 dB e Sound Pressure Lp lt 70 dB A At Operator Position e Normal Operation e According to ISO 7779 1988 EN 27779 1991 Type Test Agilent 2200 TapeStation User Manual 75 6 Appendix The Waste Electrical and Electronic Equipment Directive Abstract The Waste Electrical and Electronic Equipment WEEE Directive 2002 96 EC adopted by EU Commission on 13 February 2003 is introducing producer responsibility on all electric and electronic appliances starting with 13 August 2005 This product complies with the WEEE Directive 2002 96 EC marking requirements The affixed label indicates that you must not discard this electrical electronic product in domestic household waste Product Category With reference to the equipment types in the WEEE Directive Annex I this product is classed as a Monitoring and Control Instrumentation product Do not dispose off in domestic household waste To return unwanted products contact your local Agilent office
29. idge and tape 2 Click OK 44 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Empty Tip Buckets 1 Empty tip buckets Used loading tips must be removed from the tip buckets before the next experimental run The TapeStation will not start if tips are detected in the buckets Used ScreenTape sample strips and tips should be disposed of in accordance to local regulations How to Use the Agilent TapeStation Software For further information please refer to the software help This can be accessed by selecting the question mark button in the top right hand corner of the 2200 TapeStation Analysis Software Agilent 2200 TapeStation User Manual 45 4 46 Using the 2200 TapeStation System Shutdown and Restarting Procedure Shutdown Procedure The controller software TapeStation instrument and laptop should be shut down when not in use preferably at the end of every working day Ensure that the TapeStation System is shut down in the following order 1 Exit the TapeStation Controller Software 2 Turn off the TapeStation instrument 3 Power down the laptop Restarting Procedure Ensure that the ScreenTape System is restarted in the following order 1 Power up the laptop 2 Turn on the TapeStation 3 Start the TapeStation Controller Software Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 How to prepare your samples WARNING iat The handling of
30. l under the upper marker This can be achieved using a magnetic plate as detailed in the Sure Select protocol If you see this signal please increase the duration of this step Enlarged image of upper marker with signal from AMPure beads Figure 9 Upper marker with signal from AMPure beads Over amplification can also cause signal to run concurrently with the upper marker Agilent 2200 TapeStation User Manual 51 4 52 Using the 2200 TapeStation System Sizing Analysis software mode Within the 2200 TapeStation analysis software sizing can be found in both electropherogram and region mode The sizing methods used in electropherogram and region mode will provide different sizing information Electropherogram View Region View e Calculates data for a peak e Calculates data for a smear or region The size reported is that of the highest point Size given is that of the centre of the in the peak region s mass Lower and upper marker Always ensure that the upper and lower markers have been identified correctly The markers are used as internal references to determine the molecular weight size of the sample Incorrect identification will lead to miscalculations in reported sizing values Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation Molarity Molarity is determined from bot
31. ment will have a blue LED visible on the front of the case Agilent 2200 TapeStation User Manual Installing the System 3 4 Windows may display a Found New Hardware wizard once the software has loaded In this instance always perform the following steps a Select No not this time to prevent connecting to Windows Update and searching for software b In the next window select Install the Software automatically c Ifa window appears indicating the software did not pass the windows logo testing click Continue Anyway A window appears indicating that the hardware has been successfully installed The TapeStation system will function As there is more than one driver that can be detected and installed you may need to follow these steps more than once You may need to follow these steps if you change the USB port on the laptop for the TapeStation connector cable Agilent 2200 TapeStation User Manual 29 3 Installing the System Installing the System 30 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual 4 Using the 2200 TapeStation System Intended Use of the 2200 TapeStation System 32 Performance Limitations of Use 33 Additional Components Required by the User 34 Operating Procedure 35 Add Experiment Notes 40 Describe Samples 41 Start the TapeStation Run 43 Final Check 43 Running System 44 Abort the TapeStation Run 44 Complete TapeStation Run 44 Empty Tip Buckets 45 How to Use the Agile
32. nalysis to be saved as Do not include a full stop in file names Final Check ScreenTapeController TapeStation Controller 1 Please ensure tips and samples are loaded close the lid and click OK to continue 1 Lift the lid of the TapeStation 2 Ensure that there are fresh tips in the tip holder and that all the samples have been correctly loaded with lids removed and correspond to the sample selection on the screen 3 Close the lid Lifting the lid of the TapeStation after this time will abort the experimental run Agilent 2200 TapeStation User Manual 43 4 Using the 2200 TapeStation System Running System WARNING Exposure to potentially dangerous mechanical parts gt Do not open the lid whilst the light is flashing Abort the TapeStation Run 1 If for any reason you wish to abort an experiment click the abort button on the pop up controller The instrument will ask a If you want to reset the instrument to begin another experiment this will return the controller software and TapeStation to the beginning of the next experiment b If you want to close down the controller this will close the controller software and keep the TapeStation temporarily locked in its current state Aborting the experiment will irretrievably discard any progress made and samples loaded Complete TapeStation Run When finished a pop up will ask for removal of the tip cartridge and tape 1 Remove tip cartr
33. ng pL Lysozyme 12 5 ng pL IgG 2 uL 16 samples lt 15 min Samples consumable 16 Kit Size 112 Samples Kit Stability 4 months 1 for ladder 2 signal noise ratio gt 3 Agilent 2200 TapeStation User Manual 19 2 Specifications ScreenTape Specifications 20 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual ee 3 r Installing the System Unpacking the System 22 Contents of the ScreenTape System 24 Installing the System 27 Software Installation 27 Agilent 2200 TapeStation SetUp 28 This chapter gives information about how to install the system Apg Agilent Technologies 21 3 Installing the System Unpacking the System Unpacking the Agilent 2200 TapeStation Prerequisites Do not attempt to unpack the 2200 TapeStation instrument until you have read the accompanying Site and Safety Manual CAUTION Condensation within the instrument Condensation will damage the system electronics gt If your instrument was shipped in cold weather leave it in its box and allow it to warm slowly to room temperature to avoid condensation CAUTION Defective on arrival problems If there are signs of damage please do not attempt to install the instrument Inspection by Agilent is required to evaluate if the instrument is in good working condition gt Notify your local Agilent Representative and the Technical support channel gt An Agilent service representative will inspect
34. nt TapeStation Software 45 Shutdown and Restarting Procedure 46 How to prepare yoursamples 47 Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation 49 Quantification 49 Sizing 52 DNA Sample Preparation 54 RNA Sample Preparation 58 Protein Sample Preparation 62 This chapter explains the intended use of the 2200 TapeStation System oh Agilent Technologies 31 4 Using the 2200 TapeStation System Intended Use of the 2200 TapeStation System The 2200 TapeStation system Agilent 2200 TapeStation Software carries out electrophoretic separation of Nucleic Acids and proteins The system detects e Fluorescently stained double stranded DNA including genomic DNA e Fluorescently stained total RNA Fluorescently labelled proteins 32 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Performance Limitations of Use The 2200 TapeStation System Agilent 2200 TapeStation Software can analyse a maximum of 16 samples at any one time more samples can be run using a 96 well plate and multiple ScreenTape The user is responsible for establishing performance characteristics necessary for upstream and downstream applications Appropriate controls must be included in any upstream application requiring analysis on the 2200 TapeStation System Agilent 2200 TapeStation Software Agilent 2200 TapeStation User Manual 33 4 Using the 2200 TapeStation System Additional Components Required by th
35. ogies 2011 2012 Printed in Germany 11 2012 G2964 90001 EE Agilent Technologies
36. or see www agilent com for more information 76 Agilent 2200 TapeStation User Manual Appendix 6 Technical Service For more information please contact Agilent Technologies UK Limited e www agilent com genomics contact Agilent 2200 TapeStation User Manual 77 Index Index A analysis mode select 35 C change needle 69 check final 43 condensation 22 D delivery checklist 23 E electronic waste 76 L limitations 33 load screentape 35 tips 35 needle change 69 notes add 40 P parts damaged 23 missing 23 78 protocol pipetting 49 sample and buffer 49 ScreenTape type 49 Q quantification 49 S samples describe 41 select samples 35 test 35 service technical 77 set up TapeStation 28 shutdown procedure 46 software startup 35 sound emission 75 specifications 11 ScreenTape 14 T TapeStation abort 44 complete 44 setup 28 start 43 unpacking 22 use 35 technical service 77 tip buckets empty 45 U unpacking 22 user change 35 use TapeStation 35 wW waste electrical and electronic equipment 76 WEEE directive 76 Agilent 2200 TapeStation User Manual Appendix 6 Index Agilent 2200 TapeStation User Manual 79 www agilent com In This Book The manual describes the following Introduction to the system e Site requirements and specifications e Installation Using the system e Maintenance Product notices Agilent Technol
37. or successful loading the sample solution must be placed at the bottom of the tube or well without any air bubbles The 2200 TapeStation will load a sample from a minimum of 3 uL onto ScreenTape For best sizing precision and accuracy the user should run the appropriate ladder with the samples If 16 samples need to be analysed in parallel the user may choose to insert a saved ladder in the 2200 TapeStation analysis software Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sample Preparation High Sensitivity D1K Assay Parts required p n Description 5067 5364 High Sensitivity D1K Reagents Ladder and Sample Buffer 1 Prepare Ladder a Aliquot a minimum of 3 uL High Sensitivity D1K Ladder _ into the first tube well 2 Mix 2 uL High Sensitivity DIK Sample Buffer with 2 uL DNA sample by vortex for 5 s 3 Spin down to position the sample at the bottom of the tube 4 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section mi 2 uL HS D1K if Centrifuge C 2 uL sample 2 uL sample 2 uL sample buffer A Agilent 2200 TapeStation User Manual 55 4 Using the 2200 TapeStation System DNA Sa iple Prey aration Sample Preparation D1K Assay Parts required p n Description 5067 5362 D1K Reagents Ladder and Sample Buffer 1 Prepare Ladder a Aliquot a minimum of 3 uL D1K Ladder _ into the first tube well 2 Mix 3 uL DIK Sampl
38. otes a hazard It calls attention to an operating procedure practice or the like that if not correctly per formed or adhered to could result in damage to the product or loss of important data Do not proceed beyond a CAUTION notice until the indicated condi tions are fully understood and met A WARNING notice denotes a hazard It calls attention to an operating procedure practice or the like that if not correctly performed or adhered to could result in personal injury or death Do not proceed beyond a WARNING notice until the indi cated conditions are fully under stood and met Agilent 2200 TapeStation User Manual Contents Contents 1 Introduction to the 2200 TapeStation System 5 Overview of the System 6 2 Specifications 11 Technical Specifications 12 ScreenTape Specifications 14 3 Installing the System 21 Unpacking the System 22 Contents of the ScreenTape System 24 Installing the System 27 4 Using the 2200 TapeStation System 31 Intended Use of the 2200 TapeStation System 32 Performance Limitations of Use 33 Additional Components Required by the User 34 Operating Procedure 35 How to prepare your samples 47 Good Measurement Practices for Analysing DNA on the Agilent 2200 TapeStation 49 DNA Sample Preparation 54 RNA Sample Preparation 58 Protein Sample Preparation 62 5 Maintenance 67 General Information 68 Changing the Needle 69 Agilent 2200 TapeStation User Manual Content
39. ple degradation gt Ensure all working areas reagents and plastic ware are RNase free gt Handle RNA samples with care gt Wear gloves at all times gt Thaw RNA samples on ice gt Store RNA samples on ice throughout the ScreenTape analysis procedure CAUTION Solidification of DMSO R6K Sample Buffer contains DMSO which may solidify when cold for example if taken directly from the fridge or stored on ice gt Ensure R6K Sample Buffer is equlibrated to room temperature and mixed thoroughly prior to use gt Maintain Sample Buffer vials at room temperature throughout sample preparation gt Sample Buffer mixed with sample should always be kept on ice during sample preparation and after sample denaturation gt The Sample Buffers should be returned to 2 8 C storage once the analysis procedure has been completed 58 e Itis important to place the samples on ice directly after the denaturation step as this aids complete and stable denaturation of the RNA To ensure optimal performance of the ScreenTape R6K platform samples should be analysed using the 2200 TapeStation within 3 h of the denaturation step when left on the 2200 TapeStation system Beyond 3 h denatured samples should be stored on ice or in a suitable freezable sample block Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Do not vortex mix samples vigorously as this may degrade them When pipetting S
40. rted into the tip holder The laptop utilised for performing any previous use s of the ScreenTape must be utilised for all further re use CAUTION Damage to the 2200 TapeStation and impact on performance gt Use correct tips 4 Remove ScreenTape from the foil packet 36 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 5 Hold the tape with the ScreenTape label facing you and gently flick the top of the tape If there are any small bubbles present then this will move them to the top of the chamber The presence of small bubbles within the buffer chamber of the ScreenTape is normal These bubbles often occur at the gel buffer interface and need to be displaced prior to running Failure to remove bubbles from the gel buffer interface is detrimental to the performance of the ScreenTape 6 Insert the ScreenTape into the TapeStation with the label towards the front of the instrument and the barcode facing right Protect the individual gel lanes within the ScreenTape from excessive force Do not bend or flex ScreenTape and store in the provided packaging at the recommended temperature when not in use TapeStation instrument will not recognize the screen tape if inserted incorrectly The TapeStation will automatically recognise the sample plate type and ScreenTape and load the required parameters Agilent 2200 TapeStation User Manual 37 4 Using the 2200 TapeStation System CAUTION 7
41. s 6 Appendix 73 Limited Use Label License 74 Sound Emission 75 The Waste Electrical and Electronic Equipment Directive 76 Technical Service 77 4 Agilent 2200 TapeStation User Manual Agilent 2200 TapeStation User Manual ee 1 r Introduction to the 2200 TapeStation e System Overview of the System 6 ScreenTape Architecture 7 Agilent 2200 TapeStation Components 8 This chapter gives an introduction to the system ott Agilent Technologies 1 Introduction to the 2200 TapeStation System Overview of the System The Agilent 2200 TapeStation system is a revolutionary tape based platform for simpler faster and more reliable electrophoresis It is made up of three elements a consumable tape ScreenTape an instrument the 2200 TapeStation and an analysis software The system is very straightforward to use simply place the sample tubes and ScreenTape in to the 2200 TapeStation and let it load separate image analyse and present the results Place ScreenTape and some tips in the 2200 TapeStation Place your samples in the 2200 TapeStation and click Start on the controller software View your analysed results in around 1 min per sample This User Manual guides the operation of ScreenTape the 2200 TapeStation and software for the analysis of DNA RNA and protein samples The contents of the ScreenTape System are detailed below Information pertaining to the 2200 TapeStation can be found in
42. subsidiary of Invitrogen Corporation and Agilent Technologies UK Limited and the manufacture use sale or import of this product is subject to one or more of U S Patents and corresponding international equivalents owned by Molecular Probes Inc The purchase of this product conveys to the buyer the non transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer in conjunction with Agilent Technologies UK Limited s automated gel electrophoresis system where such research does not include testing analysis or screening services for any third party in return for compensation on a per test basis The buyer cannot sell or otherwise transfer a this product b its components or c materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes Commercial Purposes means any activity by a party for consideration and may include but is not limited to 1 use of the product or its components in manufacturing 2 use of the product or its components to provide a service information or data 3 use of the product or its components for therapeutic diagnostic or prophylactic purposes or 4 resale of the product or its components whether or not such product or its components are resold for use in research For information on purchasing a license to t
43. tion with the furnishing use or perfor mance of this document or of any information contained herein Should Agilent and the user have a separate written agreement with warranty terms covering the material in this document that conflict with these terms the warranty terms in the sep arate agreement shall control Technology Licenses The hardware and or software described in this document are furnished under a license and may be used or copied only in accor dance with the terms of such license Restricted Rights Legend If software is for use in the performance of a U S Government prime contract or subcon tract Software is delivered and licensed as Commercial computer software as defined in DFAR 252 227 7014 June 1995 or as a commercial item as defined in FAR 2 101 a or as Restricted computer soft ware as defined in FAR 52 227 19 June 1987 or any equivalent agency regulation or contract clause Use duplication or dis closure of Software is subject to Agilent Technologies standard commercial license terms and non DOD Departments and Agencies of the U S Government will receive no greater than Restricted Rights as defined in FAR 52 227 19 c 1 2 June 1987 U S Government users will receive no greater than Limited Rights as defined in FAR 52 227 14 June 1987 or DFAR 252 227 7015 b 2 November 1995 as applicable in any technical data Safety Notices CAUTION A CAUTION notice den
44. y Tw 1 uL sample buffer oT Beaten 2 uL sample k Vai 60 Agilent 2200 TapeStation User Manual Using the 2200 TapeStation System 4 Sample Preparation R6K Assay Parts required p n Description 5067 5368 R6K Reagents 1 Mix 4uL R6K Sample Buffer with 1 uL RNA sample 2 Sample denaturation Heat the samples to 72 C for 3 min Place samples on ice for 2 min Briefly centrifuge the samples to collect the contents in the base of the tubes 3 Load sample plate tips ScreenTape and Samples into the TapeStation as detailed in operating procedure section ge ane 4 uL sample buffer J geumsn Ice ioy 2 min 1 pL sample na Agilent 2200 TapeStation User Manual 61 4 Using the 2200 TapeStation System Protein Sample Preparation 62 Information on Working with Samples e When pipetting sample buffer ensure that excess buffer droplets are removed from the tip before transfer to the sample tubes Care must be taken due to viscosity of Sample Buffers e When pipetting small volumes ensure that no sample remains within the tip e When adding sample buffer to sample please ensure that they are mixed correctly To achieve this mix several times with additional pipetting then cap the tubes mix the samples using a vortex mixer on maximum speed for 5 s and briefly centrifuge to collect the contents at the base of the tubes Improper mixing can lead to quantification errors e For best results ensure that

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