Overview
Here, the DNA-RNA hybrids synthesized in Stage 1 are converted into full-length double-stranded cDNAs. The primers for synthesis of second-strand cDNA are created by RNase H, which introduces nicks into the RNA moiety of the cDNA-mRNA hybrids. E. coli DNA polymerase I extends the newly created 3'-hydroxyl termini, using the first-strand cDNA as a template. In this way, the remaining segments of mRNA in the cDNA-mRNA hybrid are replaced with the newly synthesized second strand of DNA. By Joseph Sambrook and David W. Russell.
Details
Details of this protocol, Construction Of CDNA Libraries - Stage 2: Second-Strand Synthesis (Subscription Required), are located on a web site other than Biocompare Protocols.