Transformation

Question 1

What is transformation?

Answer 1

the introduction of recombinant DNA molecules into host organisms.

Question 2

What is the common method of introducing DNA molecules into host organisms?

Answer 2

by treating E.coli with CaCl2 that partially disrupts the cell membrane (permeable) allowing DNA molecules to enter.

Question 3

What are competent cells?

Answer 3

cells that are able to take up added DNA.

Question 4

Which technique will be used to select transformants?

Answer 4

alpha complementation

Question 5

What results in blue colonies?

Answer 5

cells transformed with cloning vectors not containing inserts. (beta-galactosidase is active).

Question 6

What results in white colonies?

Answer 6

cells transformed with recombinants (beta-galactosidase gene disrupted by insert).

Question 7

Calculation of Transformation Efficiency

Answer 7

Number of transformants (microgram of DNA) * Final volume at recovery (mL) = number of transformants per microgram.