First Strand Synthesis

For each reaction required, add the appropriate multiples of the following into a master mix:

Ingredient

Volume per sample

5X First-strand buffer

2 |L

dNTPs (10 mM)

2 |L

DTT (100 mM)

1 |L

RNasin (40 U/| L)

0.05 | L

M-MLV RT (200 U/|L)

0.5 | L

H2O

4.45 | L

Dispense 10 |L of this master mix into each tube. Incubate plates in a PCR machine at 37°C (see Note 6) for 2 h for RT, 95°C for 5 min to stop the reaction and a final 4°C hold.

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