1. Requires a high-quality mRNA prep
2. Complementary strand synthesis is carried out by reverse transcriptase. Usually first-strand synthesis employs an oligo-dT primer or a primer consisting of a linearized vector with oligo-dT tails. There are many methods for second strand synthesis. Please consult methods manuals.
3. The final blunt-ended or semi-blunt-ended double-stranded cDNA can have synthetic linkers, adapters or tails added to its ends and be cloned in suitable vectors just like any other DNA.
This page was updated August 30, 1998.
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