Prepare sample. RNA serves as the template in cDNA synthesis. … Remove genomic DNA. Trace amounts of genomic DNA (gDNA) may be co-purified with RNA. … Select reverse transcriptase. … Prepare reaction mix. … Perform cDNA synthesis. … Prepare sample. … Remove genomic DNA. … Select reverse transcriptase.

How is cDNA made?

The synthesis of DNA from an RNA template, via reverse transcription, produces complementary DNA (cDNA). … Alternatively, the first-strand cDNA can be made double-stranded using DNA Polymerase I and DNA Ligase. These reaction products can be used for direct cloning without amplification.

What is used to make cDNA?

In genetics, complementary DNA (cDNA) is DNA synthesized from a single-stranded RNA (e.g., messenger RNA (mRNA) or microRNA (miRNA)) template in a reaction catalyzed by the enzyme reverse transcriptase. cDNA is often used to clone eukaryotic genes in prokaryotes.