2021 RT & qPCR

Submitted by kdorfman on Fri, 04/16/2021 - 14:09

RT

6 RNA extractions, 6 cDNA reactions/group; 23 groups

  • 65C (thermocycler)
  • pcr strips
  • ice

Give each group enough for 7 reactions

Reagent uL per rxn uL per 7 rxns
50 uM oligo(dT) 1 7
RNA
10 mM dNTP 1 7
H2O 11 71.5
5X 1st strand buffer 4 28
0.1M DTT 1 7
RNAse out 1 7
Superscript III 1 7

qPCR

48 q pcr reactions/group (24 GOI, 24 HK) Give them enough for 50 reactions

Reagent uL per rxn uL per 50 rxns
2x SYBR Green master mix 5 uL 250 uL
water 5 uL 250 uL
forward primer 0.5 uL
reverse primer 0.5 uL
cDNA 1 uL