collect RNA

Eukaryotic cell collect RNA

Eukaryotic cell transfer into E. coli

transcriptase purify DNA

ligate into plasmid

sequence ESTs

A cDNA (complementary DNA) library begins with all the RNA expressed by a cell under a specific condition. The RNA is reverse transcribed into DNA, and then each DNA copy is inserted into a separate plasmid. These are taken up by E.coli bacteria, one to a cell, which are then grown in culture. At that point, the DNA can be purified for analysis or further cloning, "expressed sequence tags" for each can be sequenced, or the proteins can be re-expressed by the bacteria to yield enough protein for further study.

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