User Manual for the 96
Contents
1. hex wrench to unscrew each of the two screws at the ends of the needle plate See Figure 6 on page 8 Keep handle assembly upside down for ease of re assembly Pull needle plate free from handle assembly Warning It may require some force to separate the seal plate from the needle plate Exercise care in handling needle plate Needles are sharp and can present a puncture hazard Also take care to avoid bending the needles Place needle plate in wash stand to stabilize it See Figure 14 Use a 3 32 hex Allen wrench to remove the 12 screws that secure the seal plate to the needle plate Revision 1 TRNDV96_MAN DOC Page 12 of 18 Figure 14 Needle plate in wash stand with all but 2 screws removed 7 Lift off the seal plate and inspect the gasket See Figure 15 Figure 15 Transfer device with plunger plate and seal plate removed and gasket exposed for inspection 8 Remove the gasket 9 Clean the seal plate and needle plate contact surfaces thoroughly with water and dry with lint free paper towels Caution Do not scratch these surfaces or use abrasives on any sealing surface 10 Make certain that all needles and wells are clean and free of debris Clean needles with cleaning wire if needed See Procedure for Cleaning Needles on page 11 11 Place new gasket on the clean dry surface of needle plate 12 Important Align the 12 hole pattern of the gasket with the pattern in the needle plate as precisely as p2. 12 replacement needles Revision 1 TRNDV96_MAN DOC Page 1 of 18 Safety The stainless steel needles used in this device may pose a puncture hazard Keep hands and fingers clear of the needles to avoid breaking skin with the needles When device is not in use it should be placed in the wash stand to protect the needles Designed for use with liquids do not use with highly viscous liquids volatile liquids or oil Do not subject the gasket to liquid when it is exposed Do not autoclave unit Use only needle cleaning wires provided for this device Revision 1 TRNDV96_MAN DOC Page 2 of 18 Using the Transfer Device There are three key steps in use of the transfer device 1 Washing the needles of the device 2 Loading of reagent 3 Dispensing of reagent into 96 wells or tubes Before using of the transfer device for the first time wash the unit with water as described below Washing the Device The wash step should be repeated at least five times as outlined in the procedure below to ensure that all reagent has been removed from the needles It is very important to wash the unit thoroughly between patient samples or between reagents to avoid cross contamination Even a minute amount of DNA material left in the unit can compromise the results of a DNA typing Procedure for Washing the Unit Figure 2 Washing the needles 1 Place a container of clean water minimum of 3 8 deep in wash stand and p
3. cceceeeeeeeeeeeeeeeeeeeeeeeneeeeeeees 12 Figure 14 Needle plate in wash stand with all but 2 screws removed 08 13 Figure 15 Transfer device with plunger plate and seal plate removed and gasket exposed for INSPECTION ccceeceeceeeeeeeeeeeeeeaeeeeeeeteeeaeeeeeeeeeeeaaeeeeeeeteesineeeeeeess 13 Figure 16 Hex wrench in vertical position ceeceeeceeeeeeeeeeeeeeeeeceeeeeeeteeetaeeeeeeees 14 Figure 17 Hex wrench in horizontal position 14 Figure 18 Screw replacement pattern for seal plate 14 Figure 19 Calibration screw on transfer dO Vcera 15 Getting Started Unpacking the Device Carefully remove the 96 Well Transfer Device from the shipping box Important Retain the shipping box and packing materials in case you need to return the unit to One Lambda for service Description of the Device Right and Left Actuators Handle Assembly Calibration Screw Plunger Plate Assembly engraved on unit in Figure 1 Transfer device stored in wash stand Product components Accessories Included e 96 Well Transfer Device Stand Catalog TRNDVSTD e Needle Replacement Tool Catalog TRNDVNDTL e Maintenance Kit Catalog TRNDVMKkIT may be ordered separately for replacement parts Accessories Not Included e Transfer Device Alignment Plate Catalog TRNDVAP required for use with the Micro SSP Gel System e Transfer Device Needle Replacement Kit Catalog TRNDVNKIT
4. Using a clean calibration syringe aspirate one well at a time in each row random column to check if the volume is within the calibration range 9 6 ul 14 4 ul Revision 1 TRNDV96_MAN DOC Page 15 of 18 3 Ifthe average volume is a value outside of the range 11 5 ul 12 5 ul adjust the LEFT calibration screw to correct the volume up to decrease and down to increase the volume 4 Repeat step 3 until the average volume is within the specified tolerance 11 5 ul 12 5 ul 5 To begin recording values on the Calibration Grid page 17 make additional copies as needed dot a new series of 96 wells Sample and record only two rows at a time 6 Using a clean calibration syringe aspirate one well at a time across the first row Record the volume starting at well 1H 1G 1F through 1A Continue this process for the second row 2H 2G 2F and so forth across the row 7 Dot a new series of 96 wells and repeat step 6 until all rows are complete 8 When you have recorded information for each well compute an average and record this value Verify that the average volume for all wells is within the calibration range 11 5 wl 12 5 ul The following steps are completed with the RIGHT wash actuator It is not necessary to record values for the RIGHT actuator unless desired or required by your laboratory s protocol 1 Depress the actuator and submerge needles in water reservoir Release the actuator to absorb water
5. 0101010101 01010101010101 0101010 000000000000000 OL J01 JOI JOI JOI J01 JOI 101 000000000000000 OX JOI JOI J01 JOI J0 JOL J01 0000000000000000 OWDOOOOOCOOOCOO0O0O 0000000000000000 OX 105 J01 101 J01 X01 101 101 000000000000000 OX J01 JOI IOI JOI J01 101 J01 000000000000000 OOOOOOCOOOOO0COVO 00000008080000008008 OOOOOOO OOOO OO0O0O 0000 00080000808008 Figure 4 Format of Micro SSP Class and Il ABDR Tray 384 SSPABDRX Procedure for Loading Samples from a 96 Well or 384 Well PCR Plate onto a 96 Well Gel 1 Before transferring a sample to the gel orient Transfer Device Alignment Plate Catalog TRNDVAP over solidified gel 2 Wash Transfer Device 5 times with clean water by pressing and releasing RIGHT actuator repeatedly and then dry needles by gently blotting them on a paper towel and repeatedly pressing the RIGHT actuator to expel all water See Procedure for Washing the Unit on page 3 3 For a 384 well tray place post PCR Well Identification Template Catalog 384TMP on the plate This assists you in transferring the correct sample onto the gel Revision 1 TRNDV96_MAN DOC Page 5 of 18 10 11 12 13 Press and hold the LEFT actuator of the transfer device before placing needles in tubes of the plate While continuing to hold the LEFT actuator down place needles in holes of template so that each needle tip touches the bottom of the tube Slowly release LEFT actuator to aspirate samp
6. Then depress the actuator again to dot a series of 96 wells on a dry hydrophobic surface such as a serum bag 2 Using a clean calibration syringe aspirate one well at a time in each row random column to check if the volume is within the calibration range 12 8 ul 19 2 ul 3 If the average volume is a value outside of the range 16 ul 20 adjust the RIGHT calibration screw to correct the volume up to decrease and down to increase the volume 4 Repeat step 3 until the volume is within the specified tolerance Troubleshooting Before calling One Lambda for assistance with your transfer device there are some simple steps you can take to resolve problems with the unit Please refer to the following table for some common problems and their solutions Problem Cause Solution Needle clogged with Too much agarose Do not use more than agarose gel 30 ml of agarose gel in the gel box To remove gel from needles soak needles in warm water and let the gel run out of the needles Then proceed with normal needle cleaning procedure Actuator was released To remove gel from before needles were needles soak needles fully removed from in warm water and let alignment plate over the gel run out of the the gel and agarose needles Then Revision 1 TRNDV96_MAN DOC Page 16 of 18 was aspirated into the needles proceed with normal needle cleaning procedure Not enough reagent in well o
7. To remove gel from needles soak needles in warm water and let the gel run out of the needles Then proceed with normal needle cleaning procedure Transfer device actuator levers stick or become stiff Loosen the two end top screws on the handle Depress the actuators until smooth and tighten screws again Revision 1 TRNDV96_MAN DOC Page 17 of 18 Calibration Grid Device Serial Number Date of calibration Technician HGFEDCBA NYL ODMYAMKWN _ Average amount dispensed Revision 1 TRNDV96_MAN DOC Page 18 of 18
8. User Manual for the 96 Well Transfer Device Catalog TRNDV96 YS One Lambda Inc Advancing Transplant Diagnostics 21001 Kittridge Street Canoga Park CA 91303 2801 Tel 818 702 0042 Fax 818 702 6904 Revision 1 Web site www onelambda com A color version of this document is available for viewing or printing at the One Lambda Web Site www onelambda com prod micro trandev index html Copyright 2000 One Lambda Inc Table of Contents GettiNG Stared irisean re raa Neea AAA AANE VEEE Geeta acs 1 Unpacking the Device Description of the Device Product components is aa ri e Acc ssones Inel uded sassa ra aa raV a a AE Washing the DeViCeis sinisa iita ti iie ee keh elated Procedure for Washing the Unit esssesseesessssiiesessrnrssstrirstststrrnttsrrrnrnsnterrnnnent Loading Liquid with the Device cccceeseeceeeeeeeeceeeeeeeeeeeeeeeeeceaeeeeeetecnaeeeeeeeeeaees Procedure for Loading Liquid Dispensing Liquid with the Device es Procedure for Dispensing Liquid eeeeeeeeeeeees Using the Device with a 96 Well or 384 Well PCR Plate DOPVICE PRE EEE EE He caidaden canduach choi A EA Mhcudtaa daweedin ceouddaadanladia taeddthalaweeduacee 7 Replacement Partsi i 28 cvnctinds ace e eal ina 7 Instructions for Return of the Device for Service 0 ee eee eect ee teneeeeeneeeeeaees 7 Replacing a Needle s 8 ofescceewcevertee hep escaeeec
9. cement parts e Transfer Device Needle Replacement Kit Catalog TRNDVNKIT 12 replacement needles Instructions for Return of the Device for Service Contact your One Lambda representative for instructions on returning the device for repairs or service You will be asked to re pack the unit in the original packing materials to ensure that the unit is properly protected for shipping Replacing a Needle The transfer device is designed for replacement of needles You should remove and replace any needle that is damaged to ensure correct operation of the device Procedure for Replacing a Needle You will need the following items for needle replacement Replacement needle s for replacement needles order the complete Maintenance Kit Cat TRNDVMKIT or the Needle Replacement Kit Cat TRNDVNKIT See above Needle replacement tool Pliers Hammer or mallet Figure 5 Replacement needles and screws are provided in a plastic tube Warning Exercise care in handling of needles Secure the needle plate before beginning the replacement process Needles are very sharp and can present a puncture hazard Revision 1 TRNDV96_MAN DOC Page 7 of 18 1 Place the transfer device upside down in the wash stand 2 To remove the needle plate from the transfer device use a 9 64 hex wrench to unscrew each of two screws at the ends of the needle plate See Figure 6 Figure 6 Transfer device placed in wash stand for rem
10. e eben Riese eiaa aaa iaa ctv uate Ead Procedure for Replacing a Needle Procedure for Cleaning Needles Gasket Replacement maun cs a Saal aTa a EE Steady Procedure for Replacing a Gasket ccccccceseeeeeeeeneeeeeeeeeeeeeeeseseeeeeeeeeeaaees Calibrating the Transfer Device 00 Transfer Device Calibration Procedure 15 Troubleshooting 16 Calibration Gid nAn E a rae at t ee Eneda act toy uate 18 Table of Figures Figure 1 Transfer device stored in wash stand Figure 2 Washing the needles ots ts Figure 3 Loading liquid 2 4 25 aig es eon i aasin in aa ran ech aAA NEEE iie Figure 4 Format of Micro SSP Class and II ABDR Tray 384 SSPABDRX Figure 5 Replacement needles and screws are provided in a plastic tube 7 Figure 6 Transfer device placed in wash stand for removal of SCreWS 0 e Figure 7 Carefully remove needle from needle plate ccceeeeeeee Figure 8 Needle replacement tool with needle inserted notched end first Figure 9 Detail of needle inserted in needle replacement tool notched end first 9 Figure 10 Seating a needle with the needle replacement tool and a mallet Figure 11 Needle plate with all needles installed c eceeseeeeeeeeees Figure 12 Placement of six springs Shown circled cccceseeeeeeeeeeeteeetteeeeeees 10 Figure 13 Illustration of Replacement Gasket
11. lace on a stable work surface A pipette tip set box or cover may be used as the water container 2 Place transfer device in stand Needles should be submerged Caution Avoid jamming of needles in reservoir 3 Press and release the RIGHT actuator at least five times to aspirate and dispense water Remove from stand 4 Press RIGHT actuator repeatedly to ensure water is expelled from the needles 5 Dry needle tips by gently blotting them on a paper towel Revision 1 TRNDV96_MAN DOC Page 3 of 18 Loading Liquid with the Device Liquid is loaded from a plate or reservoir into the unit for transfer to a 96 well format gel or plate Procedure for Loading Liquid i 2 3 lia x _ Figure 3 Loading liquid Warning Do not use the unit with highly viscous or volatile liquids Position unit over plate or reservoir from which liquid is to be loaded Press and hold LEFT actuator before touching needles to liquid Slowly release LEFT actuator to aspirate liquid into the needles Dispensing Liquid with the Device Liquid is dispensed onto a 96 well format gel or plate Procedure for Dispensing Liquid 1 For accurate placement of liquid on a One Lambda Gel System Catalog MGS 108 gel plate we recommend using a Transfer Device Alignment Plate Catalog TRNDVAP If you use the plate align it over the gel prior to loading the transfer device 2 Bring transfer device which has been loaded to the gel or pla
12. le into the needles Bring transfer device loaded with sample to the alignment plate over the gel if you are using a Micro SSP Gel System Place needles into holes of the alignment plate to ensure correct placement of the sample on the gel Press and hold LEFT actuator to dispense sample from needles onto gel Do not release LEFT actuator While continuing to hold the LEFT actuator in the down position move the transfer device and alignment plate away from the gel Check gel to ensure that the sample has been loaded in all wells of the gel The gel is ready for electrophoresis If sample loading is not complete return the alignment plate to the gel box and re insert the transfer device Continue to unload samples onto the gel Note Do not use more than 30 ml agarose gel in the Micro SSP Gel System Too much gel may clog the needles of the transfer device See Troubleshooting on page 16 if needles become clogged with agarose gel Be sure to complete Step 2 to wash the transfer device before using it to load samples from another PCR plate to a gel For a 384 well repeat steps 1 through 11 until each sample has been loaded on a gel For example for the SSPABDRX which is a four test tray you repeat these steps four times one time for each sample Revision 1 TRNDV96_MAN DOC Page 6 of 18 Service Replacement Parts The Maintenance Kit Catalog TRNDVMKIT may be ordered so that the components may be used as repla
13. ment needle cleaning wires 3 Slide the long end of the L shaped wire into each needle and slide the wire in and out to remove any built up residue or dirt Note The wire is just slightly longer than the needle so that you cannot puncture the gasket with the wire Do not depress the actuators while cleaning the needles Do not use cleaning wires other than those provided by One Lambda for use with this product Use of an incorrect wire may puncture the gasket which will lead to failure of the unit 4 Once the desired needles are clean dispense and wash the transfer device to ensure that the transfer device is operating correctly Revision 1 TRNDV96_MAN DOC Page 11 of 18 Gasket Replacement 12PL TYP 129 THRU 12PL TYP NK bS 30 2PL RIOOL Figure 13 Illustration of Replacement Gasket Procedure for Replacing a Gasket The transfer device requires a flexible gasket illustrated in Figure 13 which must be replaced when damaged A puncture or tear in the gasket indicates the need for replacement The unit must be disassembled to replace the gasket For convenience you may replace the gasket using the instructions supplied here If you wish to have the gasket replaced by One Lambda engineering staff contact your One Lambda Representative to arrange to have the gasket replaced 1 2 Place the transfer device upside down in the wash stand To remove the needle plate from the transfer device use a 9 64
14. nt pattern for seal plate 17 Reassemble the unit With handle assembly already upside down with the six springs in place place the needle plate assembly on top of the upside down plunger plate assembly Revision 1 TRNDV96_MAN DOC Page 14 of 18 Important Be sure to line the springs up in the mating holes 18 Fasten the needle plate to the base unit with the two screws you removed in step 2 Calibrating the Transfer Device We recommend that the transfer device be calibrated every one thousand trays to ensure that the appropriate amount of reagent is being dispensed The purpose of calibration is to ensure that the average amount dispensed by the needles falls within a predefined range The tolerance of the transfer device is 20 of the target dispense volume of 12 ul or 9 6 wl 14 4 ul Calibration Screw Do Nol Auloclave Figure 19 Calibration screw on transfer device Transfer Device Calibration Procedure You will need the following items to calibrate the transfer device e 80 ul calibration syringe for example Robbins Scientific Model 1060 00 0 e Serum bag or hydrophobic surface e Water reservoir e Transfer device The following steps are completed with the LEFT dispense actuator 1 Depress the actuator and submerge needles in water reservoir Release the actuator to absorb water Then depress the actuator again to dot a series of 96 wells on a dry hydrophobic surface such as a serum bag 2
15. ossible 13 With the gasket precisely aligned on the needle plate place seal plate on the assembly with the top up 14 Place the unit so that the serial number on the front edge of the plate faces you while you are assembling the unit See Figure 1 for placement of the serial number 15 Use care to minimize horizontal motion while placing the seal plate to avoid moving and mis aligning the gasket 16 Replace screws in the seal plate using the following steps Caution Make sure that gasket is clear of each screw hole If a screw is allowed to catch on the gasket the gasket may be torn or punctured Revision 1 TRNDV96_MAN DOC Page 13 of 18 a Replace the screws in the 12 holes of the seal plate using the tightening pattern shown in Figure 18 Screws should turn easily if the gasket is properly aligned This will ensure that the gasket is tightened uniformly between the plates b Tighten the screws in the pattern shown below starting at screw one and tightening in order through screw 12 until all screw heads are flush with the seal plate c With the hex wrench in a vertical position repeat the tightening pattern to tighten screws the maximum amount Figure 16 Hex wrench in vertical position d With the hex wrench in a horizontal position repeat the tightening pattern one additional time Figure 17 Hex wrench in horizontal position 113 5 9 8 1 2 T 10 6 4 12 O O Figure 18 Screw replaceme
16. oval of screws 3 To remove the needle to be replaced grip the needle end firmly with pliers and pull with a slight twisting motion perpendicular to the needle plate See Figure 7 Caution Do not apply bending motion to needle to avoid stress and possibility of needle breakage If tension is applied in a direction perpendicular to the needle plate the needle will come free Figure 7 Carefully remove needle from needle plate 4 Note that the replacement needle has a small slit or notch at one end 5 Place the replacement needle notched end first into the channel of the needle replacement tool See Figure 9 Caution If needles are not placed notched end first into the plate they will not operate correctly Revision 1 TRNDV96_MAN DOC Page 8 of 18 Figure 8 Needle replacement tool with needle inserted notched end first Figure 9 Detail of needle inserted in needle replacement tool notched end first 6 Using the needle replacement tool place the end of the needle in the hole on the needle plate from which the old needle was removed 7 Holding the needle tool perpendicular to the needle plate carefully align the protruding end of the needle into the vacant position and push the needle perpendicular to the needle plate until the needle end of the replacement tool is flush with needle plate 8 Sufficient hand force will seat the needle with the replacement tool However if you cannot seat the needle with hand force u
17. r on plate Gasket or needle has exceeded useful life span Replace gasket and or needle Needle does not aspirate or dispense reagent Needle placed upside down un notched end out in plate Replace needle with notched end out Problem Cause Solution No reagent in well or on plate Gasket or needle has exceeded useful life span Replace gasket and or needle Reagent is dispensed unevenly in wells or on plate Gasket or needle has exceeded useful life span Spring is missing or damaged Needles do not align with wells Replace gasket and or needle Check for missing or damaged springs replace springs as needed Make sure all needles are seated at the same height in the plate make sure needle rows are aligned along both the width and length of the transfer device Needle leaks Leak in gasket Leak at the junction of the needle and the plate Replace gasket Replace needle seal needle at base with super glue Transfer device sticks to alignment plate or does not slide easily out of alignment plate User is not maintaining needles perpendicular to the alignment plate when pulling the transfer device out of the alignment plate There is gel on the needles Pull transfer device out of alignment plate ina vertical direction perpendicular to the alignment plate Do not use more than 30 ml of agarose gel in the gel box
18. se a hammer or mallet to tap on the end of the needle replacement tool until the tool is flush with the needle plate See Figure 10 Multiple light taps are preferable to a single hard blow on the replacement tool Figure 10 Seating a needle with the needle replacement tool and a mallet 9 Check each replaced needle for alignment by completing the following steps e Sight down each row of needles along the length of the needle plate e Sight down each row of needles along the width of the needle plate Revision 1 TRNDV96_MAN DOC Page 9 of 18 If a needle is out of alignment with other needles in the row apply a gentle bending hand force to that needle until it is straight and aligned with the other needles along both the length and width of the plate See Figure 11 Figure 11 Needle plate with all needles installed 10 Reassemble the unit Place the needle plate on the handle assembly Be sure to line the springs up in the mating holes See spring placement in Figure 12 below Fasten the needle plate to the base unit with the two screws you removed in step 2 Figure 12 Placement of six springs shown circled Revision 1 TRNDV96_MAN DOC Page 10 of 18 Cleaning the Needles Procedure for Cleaning Needles 1 Place transfer device upside down on wash stand or hold it upside down in your hands 2 Use a One Lambda Needle Cleaning Wire Catalog TRNDWKIT Transfer Device Needle Cleaning Wire Kit 12 replace
19. te on which the liquid is to be dispensed If you are using a transfer device alignment plate the transfer device should mate with or meet the alignment plate to ensure correct placement on the gel The needle plate should be flush with the alignment plate 3 Press and hold the LEFT actuator to dispense samples onto the gel or plate 4 While maintaining the LEFT actuator in the down position move the transfer device away from the gel or plate 5 Verify that reagent has loaded properly on the gel or plate If the reagent has not been loaded properly return the transfer device to the gel or plate and dispense remaining liquid 6 Complete steps in WASHING THE UNIT on page 3 before loading and dispensing a new reagent or sample Revision 1 TRNDV96_MAN DOC Page 4 of 18 Using the Device with a 96 Well or 384 Well PCR Plate One Lambda s 384 well Micro SSP trays contain multiple tests For example the Micro SSP ABDR HLA DNA Typing Tray Catalog SSPABDRX contains 4 tests Each test sample is loaded in a 96 well format In the example shown in Figure 4 sample 1 is loaded in all of the white wells sample 2 is loaded in all of the light gray wells sample 3 is loaded in all of the dark gray wells and sample 4 is loaded in all of the black wells COOOOCOOVOO0OD OOP 0000000008888 Sw OOOOOOVOO0OO0O 000000000000000 OQOOOOQOOOOOQOO0OQOO0OOQO0OQO 000000000000000 OF XOF ZOI 101 101 101 JOL J01 000000000000000
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