2021 RNA extraction

Submitted by kdorfman on Fri, 04/16/2021 - 13:50

RNEasy Kit Qiagen 74904

Per reaction (6 x 23=138), :

  • LN2
  • Tissue flash frozen in LN2 in round bottom 2 mL tube with 2 grinding beads
  • Buffer RLT 450 uL
  • lilac QIAshredder in 2 mL collection tube
  • pink tube
  • 225 uL 100% ethanol
  • 350 Buffer RW1 x 2 (see below)
  • 10 uL DNAse (in RDD) IF NO INTRON
  • 70 uL RDD
  • 350 uL RW1
  • 500 uL RPE
  • 500 uL RPE
  • 30 uL RNAse free water
  • 20 uL RNAse free water
Reagent uL per rxn uL per 6.5 rxns
RLT 450 2925
EtOH 225 1462.5
RW1 700 4550
DNAse 10 65 (have ~570 uL, more on the way)
RDD 70 455
RPE 1000 6500
H2O 50 325