Preparation of cDNA

1. All reactions are done in 0.2 mL PCR tubes. Mix 5 micrograms of RNA with 2 picomol of reverse primer.

2. Add H20 to make up a volume of 12 microlitres.

3. Incubate at 70 celsius for 10 minutes. Chill immediately on ice after incubation.

4. Subsequently add 4 microlitres of first strand buffer, 2 microlitre of 0.1 M DTT, 0.5 microitre dNTPs from a 100mM stock, 0.5 microlitre RNaseIn and incubate for 2 minutes at 42 celsius.

5. Add 1 microlitre SuperScript II RT.

6. Incubate