Imagine you wanted to use a human genomic DNA library to clone the human gene for insulin. You will be using the rat insulin gene sequence as your DNA probe. In what order would you perform the following steps to accomplish this goal? 1. Use autoradiography to identify colonies containing DNA that hybridized to the probe
2) Grow transformed cells on media with antibiotics and X-gal for blue-white screening
3) Ligate genomic DNA and vector DNA
4) Cut genomic DNA and vector DNA with restriction enzymes
5) Hybridize library DNA with labeled probe for the rat insulin gene
6) Transform bacteria with recombinant plasmid
A) 2, 3, 6, 5, 4, 1
B) 1, 2, 5, 6, 4, 3
C) 4, 3, 6, 2, 5, 1
D) 4, 3, 5, 2, 6, 1
E) 2, 6, 5, 3, 1, 4

Question

Quizplus · Accepted Answer

The correct order for cloning the human insulin gene using a rat insulin gene sequence as a probe involves: cutting both genomic DNA and vector DNA with restriction enzymes (4), ligating the genomic DNA into the vector (3), transforming bacteria with the recombinant DNA (6), growing the transformed cells on media for selection (2), hybridizing the library DNA with the labeled probe (5), and finally using autoradiography to identify the colonies that have hybridized with the probe (1).

Imagine You Wanted to Use a Human Genomic DNA Library