2021 RT & qPCR
2021 RT & qPCR kdorfman Fri, 04/16/2021 - 14:09RT
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 |