Question

53: WHICH technique is NOT used to make a cDNA library? A. Extraction of DNA from...

53: WHICH technique is NOT used to make a cDNA library?

A. Extraction of DNA from the organism you are studying

B. Extraction of RNA from the organism you are studying

C. Use of reverse transcriptase to copy RNA into DNA

D. Ligation of DNA into a cloning vector

E. Insertion of cloning vector into cells

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Answer #1

Process of making cDNA library:

1. First the mRNA is obtained the organism of interest and purified from the rest of the RNAs.

2. Reverse transcriptase is uses on purified mRNA to create the complementary DNA strand. The mRNA is removed using a RNAse enzyme leaving a single stranded cDNA.

3. Restriction endonucleases and DNA ligase are then used to clone the sequences into bacterial plasmids.

4. The cloned bacteria are selected, and later grown and sequenced to compile the cDNA library.

ANSWER: B) Extraction of RNA from the organism you are studying.

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