Gene Cloning

Question 1

What is a cDNA library?

Answer 1

a collection of cloned DNA sequences (vDNAs) that are complementary to the mRNA extracted from the organism/tissue.

Question 2

What is a genomic library?

Answer 2

contains cloned fragments of the entire genome of the organism

Question 3

What do you need for library screening? (2)

Answer 3

1. replica plate of the cloned cells

2. DNA probes (complementary to the sequence you are screening for)

Question 4

Role of DNA polymerase I in making a cDNA library

Answer 4

synthesises cDNA (using RNA fragments as primers)

also has endonuclease activity to degrade mRNA as it goes along

Question 5

Role of RNAseH in making a cDNA library?

Answer 5

generates a nick in RNA molecule to create RNA fragments

Question 6

How is DNA hybridisation involved in cDNA library screening?

Answer 6

Made a replica of the cloned cells

add probe with buffer

probe binds to complementary sequence of cDNA that you are looking for