User Bulletin ABI PRISM 377 DNA Sequencer
Contents
1. io 5 Gel p i temperature i control unit is Pg oe ee ae fp gr by passed CO n GJ Pump Heat HS za exchange Heater 5 N i A 0 SA Figure 2 ABI PRISM 377 interior view with arrows showing water flow Dotted line shows ABI PRISM 377 components used with the external water bath If the external water bath you use has heating capacity to 60 C you can run the instrument under standard configuration conditions ambient temperature plus 10 C to 60 C with the external bath attached Frequent monitoring of the water level is required when the external bath is used during standard runs For long periods of use disconnect the external water bath reinstall the water bottle and tubes and use the standard run modules not the chiller modules in the Data Collection 2 1 software Modules folder Refer to the Data Collection Software 2 1 Enhancements user bulletin for the names of all the Data Collection 2 1 software modules and for setting preferences to switch between the standard run modules and the chiller modules User Bulletin ABI PRISM 377 DNA Sequencer Page 5 of 10 Installing the External Cold Water Bath Positioning the To position the ABI PRISM 377 and external water bath Machines Step Action 1 Place the ABI PRISM 377 on a table or counter that provides easy rear access Facing the ABI PRISM 377 position the water bath on the right side of the instrument and at the s2. installed on your instrument upgrade to the Data Collection 2 1 software and check the Log file from a recent run The configuration information in the Log makes reference to the type of thermistors installed Schedule a service call with your PE Applied Biosystems service engineer for replacement of the 100 k ohm thermistors if you wish to run SSCP at the lower temperatures Software The Data Collection 2 1 software upgrade contains all required Requirements features to accommodate the external cold water bath including new chiller modules Refer to the user bulletin ABI PRISM 377 DNA Sequencer Data Collection Software 2 1 Enhancements P N 904945 for software installation instructions descriptions of features and instructions for using the new modules Requirements for Specific Run Thermistor External Conditions Run Conditions Software ohms Bath gt 10 C above ambient to 60 C 1 1 or 2 1 100 k not required SSCP from 22 C to ambient 2 1 100 k required 10 C SSCP at lt 22 C 2 1 30 k required User Bulletin ABI PRISM 377 DNA Sequencer Page 3 of 10 How the External Cold Water Bath Functions Disengages The ABI PRISM 377 configuration with the external cold water bath Internal System attached is similar to Figure 1 when viewed from the back New modules in the Data Collection 2 1 software disengage the instrument s internal pump system and switch to the external cold water bath When you
3. turn the external valve so the arrow points down access to the internal gel temperature control unit is closed off TO External valve HOTQUUQUOOQUOQUCAUOANNOGUNOUOOOUOOUUOUTOQUUOUNOUNOOUEOUOOOUEOIUGI arrow down mang ran f ees Le mists ge OOOIOUUOOUIOUNOOIONOOOOOONOOUTOUNJETOUNOODOUNOOTOUNDONMOUNIN TI aie AALAOUUOUOOU OODI OOUOO OONO UOU UOUU VOOU DUOOUON UON OO OOUT TOETAN AAOOOOUIIOOIIUNOIOUOOOOOOOOUUOOOONYOIA00ULOOOOOOUOOOOOOONIIUINII IIH Figure 1 Rear view of the ABI PRISM 377 with external water bath attached GR0686 Water Bypasses the Gel Temperature Control Unit With the external cold water bath attached water passes directly into the front and back gel heat transfer plates in the instrument bypassing the gel temperature control unit Figure 2 Page 4 of 10 User Bulletin ABI PRISM 377 DNA Sequencer Standard Configuration Run Conditions with External Bath Attached External Valve arrow down Flow sensor rf SS a ee ee ee ee N Xi HA H i l Ip ual CS TF Front plate t J External Back plate water Ho bath i
4. User Bulletin ABI PRISM 377 DNA Sequencer June 5 1997 SUBJECT Modifications for Subambient Temperature Operations Purpose of this This user bulletin describes how to modify your ABI PRISM 377 DNA User Bulletin Sequencer to accommodate an external gel cooling system for run temperatures below the standard temperature The standard run temperature is ambient plus 10 C up to 60 C The following topics are included Topic See page Purpose of an External Cooling System 2 Hardware and Software Requirements How the External Cold Water Bath Functions Installing the External Cold Water Bath Operating at Subambient Temperatures oloo AIN Avoiding Condensation PE Applied Biosystems A DIVISION OF PERKIN ELMER Purpose of an External Cooling System Performing SSCP The ABI PRISM 377 DNA Sequencer provides gel temperature control from 10 C above the ambient temperature to a maximum of 60 C To perform PCR Single stranded Conformation Polymorphism SSCP protocols at lower temperatures below 10 C above ambient you must attach a cold water bath to the instrument Refer to PCR SSCP Analysis A Guide to Fluorescent PCR Single stranded Conformation Polymorphism Analysis on the ABI PRism 377 DNA Sequencer P N 904413 for further information Hardware and Software Requirements Overview Hardware Requirements Page 2 of 10 To provide gel temperature control for o
5. ame height Plug both machines into an external power supply Determining To identify the intake and outlet flow tubes on the instrument Action Turn on the ABI PRISM 377 Open the right side panel where the water bottle for the internal pump system is mounted Unscrew the water bottle and slowly pull it down to expose the ends of the tubes but do not remove the bottle Observe the direction of water flow from the tubes into and out of the bottle to identify the intake and outlet tubes Note Water coming into the bottle is from the instrument outlet and water being drawn out of the bottle is pumped into the instrument intake Some instruments have a small cutout at the bottom of the water intake tube Turn off the ABI PRISM 377 Intake and Outlet Step 1 2 3 4 5 Page 6 of 10 continued on next page User Bulletin ABI PRISM 377 DNA Sequencer Disconnecting the Internal Pump Attaching the External Cold Water Bath To disconnect the instrument s internal pump Step Action 1 Remove the water bottle and pull down the intake and outlet tubes to remove them Store the tubes and water bottle for later use Step to the rear of the instrument and turn the external valve so that the arrow points down for the external position 4 Cut two pieces of polyurethane tubing approximately three feet one meter long to connect the external wate
6. perations at reduced temperatures you must have all of the following an external cold water bath connected to the ABI PRISM 377 30 k ohm thermistors installed on the instrument the ABI PRISM 377 Data Collection 2 1 software The water bath polyurethane tubing and tubing connectors needed for the instrument modification must meet the following minimum requirements Hardware Minimum Requirements Water Bath Cooling capacity 250 watts at 20 C Heating capacity 500 watts up to 60 C Pump flow rate 1 gal min or 4 L min at pressure head of 3 m Automatic Shutdown At high temperature and low liquid level Polyurethane Tubing 6 ft 1 4 in i d 3 8 inch o d Tubing Connectors 1 4 in female NPT 1 4 in male barbed fittings NESLAB P N 126000000007 User Bulletin ABI PRISM 377 DNA Sequencer Any external water bath that meets these requirements should work properly The NESLAB Model RTE111 meets these requirements and has been tested with the instrument Refer to the user manual for the water bath you select follow proper procedures and ensure that all product safety and regulatory requirements applicable to your location are met Changing Thermistors for SSCP Most ABI PRISM 377 instruments have 100 k ohm thermistors installed To perform SSCP at temperatures less than 22 C you must replace the 100 k ohm thermistors with 30 k ohm thermistors To determine which thermistors are
7. power to the external water bath start the circulating pump and add water as needed to maintain the proper level IMPORTANT If the water level drops below the level recommended by the manufacturer the circulation pump shuts off and the required temperature is not maintained 5 Set the external cold water bath temperature about 3 4 C below the desired gel plate temperature and monitor the temperature in the Status screen under the Window menu Place a lid on the bath to minimize evaporation Add water and antifreeze during operation as needed to maintain the manufacturer s recommended water level 8 Monitor the temperature and humidity in the lab refer to the following section Avoiding Condensation 9 Refer to the user bulletin ABI PRISM 377 DNA Sequencer Data Collection Software 2 1 Enhancements P N 904945 for software installation procedures Page 8 of 10 User Bulletin ABI PRISM 377 DNA Sequencer Avoiding Condensation Severe Damage May Result Operating the ABI PRISM 377 DNA Sequencer at subambient temperatures may cause condensation to form inside the instrument which can cause decreased performance and severe damage To avoid condensation you must keep the water temperature above the dew point temperature at which water vapor condenses into liquid or lower the humidity in the room Consult the following tables to determine the dew point of water at various relative humidi
8. r bath to the intake and outlet openings on the instrument 5 Insert one end of a piece of tubing into the intake port on the instrument 6 Repeat step 5 with the second piece of tubing in the outlet port To attach the external cold water bath to the instrument Step Action 1 Wrap the threads of the intake and outlet ports on the external water bath with Teflon tape 2 Attach a metal tubing connector to each port Connect the free end of the intake tube to the outlet fitting on the back of the external water bath 4 Connect the free end of the outlet tube to the intake fitting on the back of the external water bath User Bulletin ABI PRISM 377 DNA Sequencer Page 7 of 10 Operating at Subambient Temperatures Performing a Runat After your ABI PRISM 377 is modified for subambient temperature Subambient runs perform the following steps each time you use it Temperatures P To perform a run at subambient temperatures Step Action 1 Follow all instructions in the external water bath user s manual 2 Fill the external water bath with cold deionized distilled water and add antifreeze as recommended by the manufacturer WARNING CHEMICAL HAZARD Antifreeze may cause respiratory tract skin and eye irritation Wear chemical resistant gloves and safety glasses when handling and always use in a well ventilated area 3 Check that all tubing is tightly connected 4 Turn on
9. ties and ambient temperatures in your lab CAUTION Condensation can cause decreased performance and severe damage to the instrument Monitor the room temperature and humidity consult the following condensation tables and maintain the water bath temperature setting above the corresponding dew point Dew Point Table Ambient Temp Ambient Temp Ambient Temp Ambient Temp Ambient Temp 15 C 20 C 35 C Approx Approx Approx Approx Approx Relative Dew Relative Dew Relative Dew Relative Dew Relative Dew Humidity Point Humidity Point Humidity Point Humidity Point Humidity Point CC C C C C 100 15 100 20 100 25 100 30 100 35 90 14 90 18 90 23 90 29 90 33 80 12 80 16 80 21 80 26 80 31 70 10 70 14 70 19 70 24 70 29 60 12 60 17 60 21 60 26 50 10 50 14 50 19 50 23 40 10 40 15 40 19 30 10 30 15 20 9 User Bulletin ABI PRISM 377 DNA Sequencer Page 9 of 10 Copyright 1997 The Perkin Elmer Corporation Perkin Elmer is a registered trademark of The Perkin Elmer Corporation ABI ABI PRISM and the ABI PRISM design Applied Biosystems PE and PE Applied Biosystems are trademarks of The Perkin Elmer Corporation All other trademarks are the sole property of their respective owners P N 904056 Stock No 237837 001
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