Creating a cDNA library

1. Isolate mRNA from [tissue] 
2. Use reverse transcriptase to synthesise a cDNA strand using RNA as a template
3. Use DNA polymerase to make double-stranded cDNA and amplify using PCR
4. Cut plasmid with restriction enzyme that leaves blunt ends
5. Use terminal transferase to add extra cytosine nucleotides to the blunt end of DNA strands and add guanine nucleotides to that of plasmids to form sticky ends
6. Mix plasmid with cDNA; complementary sticky ends anneal to one another by hydrogen bonding;
7. Add DNA ligase to seal nicks by forming covalent phosphodiester bonds to form recombinant plasmid