Ajeffs:bioanalyzer: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(New page: Back to Eccles Lab > DGG Protocols ==Bioanalyzer pre-flight check list== ===Reagents=== #All reagents, except for the ladder, are in the top door-shelf of ...)
 
No edit summary
Line 6: Line 6:
#Reagents, except ladder, must stand at room temperature for 30 minutes before use.
#Reagents, except ladder, must stand at room temperature for 30 minutes before use.
#Check the date on the filtered gel in the reagent box: discard and make up fresh if too old (Agilent say 4 weeks, but we have used it for 2-3 times longer with no obvious loss in performance).
#Check the date on the filtered gel in the reagent box: discard and make up fresh if too old (Agilent say 4 weeks, but we have used it for 2-3 times longer with no obvious loss in performance).
#The ladders are in 0.2 mL tubes in Box 153 of the NZORD -80dC in Hercus 23x? The ladders are heat-denatured and in 1 uL aliquots. Make sure you get the right ladder: Nano should be the top rack of two inside the box, Pico the second rack and on the bottom.
===Laders===
*The RNA ladders are in 0.2 mL tubes in Box 153 of the NZORD -80dC in Hercus 23x? The ladders are heat-denatured and in 1 uL aliquots. Make sure you get the right ladder: Nano should be the top rack of two inside the box, Pico the second rack and on the bottom.
*The DNA ladders are not stored at -20dC, and are in the box with the rest of the reagents at 4dC.


===Hardware===
===Hardware===

Revision as of 17:16, 6 February 2011

Back to Eccles Lab > DGG Protocols

Bioanalyzer pre-flight check list

Reagents

  1. All reagents, except for the ladder, are in the top door-shelf of the fridge in Hercus 210.
  2. Reagents, except ladder, must stand at room temperature for 30 minutes before use.
  3. Check the date on the filtered gel in the reagent box: discard and make up fresh if too old (Agilent say 4 weeks, but we have used it for 2-3 times longer with no obvious loss in performance).

Laders

  • The RNA ladders are in 0.2 mL tubes in Box 153 of the NZORD -80dC in Hercus 23x? The ladders are heat-denatured and in 1 uL aliquots. Make sure you get the right ladder: Nano should be the top rack of two inside the box, Pico the second rack and on the bottom.
  • The DNA ladders are not stored at -20dC, and are in the box with the rest of the reagents at 4dC.

Hardware

  1. Check the settings on the priming station:
    • Position of the chip deck: A, B, C, or D.
    • Position of the syringe clip: top, middle, or bottom.

Tips

  1. Pipette gel and marker onto the bottom of the well, not the side.
  2. Follow the Agilent protocol exactly.
  3. Pipette 6 uL of marker into non-sample-containing wells during the first pass: this saves a second pass that adds 1 uL of extra marker to the first 5 uL of marker in non-sample-containing wells to make up the total volume to 6 uL.
  4. Always load sequentially from sample well number one.
  5. If running less than a full chip, reduce the run time by altering the number of samples to run on the "Instrument" page to match the actual number of samples you have, remembering that the sample order zig-zags from top-left to bottom-right.

Decontamination

  1. 350 uL RNase Zap, 1 min.
  2. 350 uL nuclease-free water, 10 sec.
  3. Air-dry, 10 sec.