Applied Biosystems 3730/3730xl DNA Analyzer

Transcription

1 pplied iosystems 0/0xl nalyzer pplied iosystems 0/0xl nalyzer Quick Reference ard This quick reference card supplements the pplied iosystems 0/0xl nalyzers etting Started uide ( ). or safety guidelines, refer to the 0/0xl nalyzers etting Started uide. RTT! Refer to the aterial Safety ata Sheets (SSs) for chemicals listed in bold type for information. Wear gloves any time you handle the capillary array, septa, buffer reservoirs, polymer bottles, polymer blocks, interconnect tubing, and bottle cap tubing.. ill the cathode buffer, water, and waste reservoirs with 0 m of the appropriate liquid.. ill the clean, dry anode buffer reservoir with m of running buffer. RTT! The meniscus should line up just under the red fill line before you install the anode buffer reservoir on the instrument (see figure below). Starting the pplied iosystems 0/ 0xl System o not rename the computer after the pplied iosystems 0/0xl nalyzer ata ollection Software has been installed. oing so may cause the 0/0xl ata ollection software to malfunction. RTT! e sure to start the computer workstation before starting the pplied iosystems 0/0xl nalyzers. Step : Start the computer workstation. ower on the monitor.. ower on the computer. The egin ogon dialog box opens.. nter the user name and password (there is no default password) for the ata ollection Software logon. Step : Start the 0/0xl nstrument ote: f the anode buffer jar buffer overflows as polymer is added, discard and then replace with fresh X running buffer. The anode buffer jar could overflow during bubble clearing. olymer locks R0 eniscus should be just under the fill line node buffer jar ssemble the lower polymer block as shown below: Wear gloves any time you handle the capillary array, septa, buffer reservoirs, polymer bottle, polymer block, interconnect tubing, and bottle cap tubing.. nsure that the computer is powered on.. n the instrument, ensure that the: ven door is closed nstrument door is closed Stacker drawer is closed uffer, water, and waste trays are loaded.. Turn on the instrument. Step : rerun Setup. ttach a clean, dry lower polymer block to the instrument.. nstall the capillary array using the nstall apillary rray wizard. RTT! When analyzing with basecaller in Sequencing nalysis software, use the apillary rray wizard to input the applicable capillary length information.. nstall the polymer bottle. See the hange olymer wizard.. repare the running buffer. ump block heck valve olymer block interconnect tubing assembly uffer pin valve ower polymer block olymer bottle cap assembly node electrode olymer bottle uffer jar RT pril, 00 : pm,.fm

2 hecking for ubbles ubbles can collect where the channels join. The following figure illustrates where bubbles may collect. Sealing lates with eat Seal ilm -Well late ssembly Using a eat-sealed late Watch for bubbles: t the array ferrule tip n all tubing late retainer eat seal film eat-sealed sample plate n the elbow (internal channel) of the lower block R ray plate base Removing ir ubbles from the olymer lock To remove air bubbles, select Wizards > ubble Remove Wizard, then follow the instructions in the wizard. ssembled components R Sealing lates with Septa Supported ye Sets pplied iosystems 0/0xl ata ollection Software v.0 supports: Septa-ased late ssembly lign the septum holes with the plate sample holes late septum pplication hemistry ye Set hemistry ile Sample plate ragment nalysis RS inkage apping Set v. -RT atrix Standard Sequencing igye Terminator v. -igyev Sequencing Standard late retainer igye Terminator v. Z-igyeV Slex Slex hemistry Suggested oading Volumes S atrix Standard late septum Sample plate ZR. ormamide causes eye, skin, and respiratory tract irritation. t is a possible reproductive and birth defect hazard. Read the SS, and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. repare the formamide:size-standard mix. or loading a: -well plate, use µ per well (minimum volume). -well plate, use µ per well (minimum volume). R lack plate base lign the notched corners RT pril, 00 : pm,.fm age

3 Run odules nalysis Type apillary rray ength (cm) Run odule ragment analysis eneapper_ xtra long-read 0 eneapper0_ 0 XRSeq0_ ong 0 ongseq0_ ast-read Standard-read Rapid-read 0 astseq0_ StdSeq_ RapidSeq_ Slex cm TS V Slex 0 cm 0 TS0_. ttach the syringe to the forward-facing uer fitting at the top of the pump block. old the fitting with one hand while threading the syringe onto the fitting with the other hand.. pen the uer fitting by grasping the body of the fitting and turning it and the attached syringe approximately one-half turn counterclockwise.. pen the exit fitting at the top left side of the pump block by turning it approximately one-half turn counterclockwise.. While holding an empty tube or beaker under the exit fitting, flush the trap at a flow rate of m per 0 sec by pushing steadily on the syringe plunger. T US XSSV R when you push the syringe plunger. xcessive force may damage the trap seals. Take approximately 0 seconds to flush m of either distilled or deionized water through the trap. art umbers for igh-use tems tem art o 0 -cm capillary array 0 0-cm capillary array 0 0xl -cm capillary array 0xl 0-cm capillary array lushing and illing the Water Trap lush the polymer delivery pump water trap at least once per week to wash out any diluted polymer that may have passed through the water trap seals. Some air bubbles in the water trap are acceptable and do not affect performance. eave the trap filled with either distilled or deionized water. ote: The figure below shows a beaker being used to catch the trap s flushed waste fluid. 00/0 igye Terminator v. Sequencing Std 00/0 igye Terminator v. Sequencing Std atrix Standard Set S- ii formamide, m 0 - ( bottle of -m) - ( bottles of -m each) - (0 bottles of -m each) uffer ( ) with T, 00 m uffer ( with T, -well sample plates w/bar code 0 -well sample plates, no bar code well plate septa -well plate base (septa sealed) -well plate base (heat sealed) -well plate retainer (septa sealed) -well and -well plate retainer (heat sealed) -well sample plates with barcode 0 -well plate septa To flush the water seal trap:. ill the supplied 0-m, all-plastic uer lock syringe ( ) with distilled or deionized water. xpel any bubbles from the syringe. ote: o not use a syringe smaller than 0 m. oing so may generate excessive pressure within the trap. -well plate base (septa-sealed) -well plate base (heat-sealed) -well plate retainer (septa-sealed) eat seal film, -mil 0 RT pril, 00 : pm,.fm age

4 nstrument aintenance Schedule RTT! Refer to the aterial Safety ata sheets (SSs) for chemicals listed in bold for safety information. Wear gloves any time you handle the capillary array, septa, buffer reservoirs, polymer bottles, polymer blocks, interconnect tubing, and bottle cap tubing. aily aintenance Task requency Weekly aintenance Task lean the: Water, waste, and buffer reservoirs with warm water and a thorough distilled/ deionized water rinse omplete polymer path including the pump and lower polymer blocks requency Weekly or when the polymer is changed Replace the appropriate liquid in the water, waste, and running buffer reservoirs on the instrument. RTT! o not top off buffer/water/ waste fluids after a run. mpty the reservoir fluids every hours and then refill with fresh solutions for optimal performance. heck for dried polymer around the polymer block and clean as necessary. very to hours aily, or before each batch of runs lush the polymer delivery pump water trap. Replace the polymer in the bottle, pump block, and capillary array. Restart the computer and instrument. heck the storage conditions of the used arrays. heck database space and hard disk space. Weekly Weekly or as needed Weekly nsure adequate: uffer and water levels in the cathode reservoirs uffer level in the anode buffer jar olymer level in the bottle for scheduled runs s eeded aintenance Task requency nsure that the: late assemblies are put together properly. The holes in the plate retainer must align with the holes in the septa; otherwise the capillary tips will be damaged. Reservoir plate assemblies are positioned on the plate deck properly. lates should sit snugly on the deck. ever use warped plates. lean the: rip tray nstrument surfaces hange the array. Remove any dried polymer from the capillary tips. Use a lint-free wipe moistened with deionized water. s needed heck for and remove all bubbles from the: ower polymer block nterconnecting tubing olymer cap tubing olymer block channels efragment the computer hard drive. s needed or monthly heck the loading-end header to ensure that the capillary tips are not crushed or damaged. heck the lower polymer block to ensure that it fits securely on the instrument. heck for leaks around all fittings on the pump and lower polymer blocks. RT pril, 00 : pm,.fm age

5 -apillaries oad ap -Well late oad ap s = irst load irst-quadrant pickup Second-quadrant pickup = irst load 0 Third-quadrant pickup = irst load = irst load 0 ourth-quadrant pickup 0 0 = irst load RT pril, 00 : pm,.fm age

6 -capillaries oad ap -Well late oad ap Well late oad ap irst-quadrant pickup Second-quadrant pickup Third-quadrant pickup ourth-quadrant pickup opyright 00, pplied iosystems. ll rights reserved. or Research Use nly. ot for use in diagnostic procedures. The pplied iosystems 0 and 0xl nalyzers include patented technology licensed from itachi, td. as part of a strategic partnership between pplied iosystems and itachi, td., as well as patented technology of pplied iosystems. RS, pplied iosystems, igye, and eneapper are registered trademarks and (esign), pplera, i-i,, and are trademarks of pplera orporation or its subsidiaries in the U.S. and/or certain other countries. ll other trademarks are the sole property of their respective owners. 0/00 RT pril, 00 : pm,.fm art umber Rev.