Bioanalyzer Samples

  1. Make sure customers know we only keep DNA 1000 chips on hand; expected bands need to be between 15 and 1500 bps.

  2. Measure concentrations of ALL SAMPLES on the Nanodrop or Synergy HTX

  3. If samples are below 1 ng/ul, reject. If samples are above 50 ng/ul, dilute to 40 ng/ul.

  4. Pull “Agilent DNA 1000 Reagents” from Fridge 5 and allow to equilibrate to RT for 30 minutes minimum.

  5. If date written atop a clear tube has not passed, proceed to step 10

  6. Add 25 ul of DNA dye concentrate (blue dot) to a DNA gel matrix vial (red dot)

  7. Vortex solution well and Centrifuge down. Transfer full volume to spin filter.

  8. Centrifuge at 2250 rcf for 15 minutes.

  9. Discard filter. Mark top with the date 6 weeks from today.

  10. Remove a DNA 1000 chip from its plastic sleeve and place on open priming station (use grey lever to release if latched)

  11. Vortex and spin the dated tube(gel-dye), the yellow tube (ladder), a green tube (markers),

  12. Pipette 9 ul of gel-dye mix into the bottom of the lower right well marked with a white G in a black circle

  13. Make sure the plunger is positioned at 1 ml. Close priming station until a click is heard.

  14. Depress plunger until it is held by the catch and start 1 minute timer

  15. Release plunger and watch it release pressure until ~0.7 ml and then slowly assist back to 1 ml

  16. Open priming station and add 9 ul gel-dye mix to the bottom of the two upper wells marked with black Gs

  17. Load 5 ul to the bottom of the well marked with a ladder symbol and all numbered wells.

  18. Load 1 ul ladder to the ladder well

  19. Load 1 ul sample or water to all numbered wells. (1 ul must be added to all sample wells in order for a chip to run successfully)

  20. Transfer the chip to the chip vortexes set at 2400 rpm and turn on for 1 minute.

  21. Open Bioanalyzer and place chip into bay. A notch built in the chip prevents improper loading.

  22. Open 2100 expert software on computer. Make sure the correct instrument is selected and there are no red Xs in the bottom right and the proper assay is displayed, “DNA 1000”. Then press “start”.