in vitro cloning

Question 1

What is the first step for in vitro cloning?

Answer 1

= temperature is first increased to 90 degrees, to break the complementary hydrigen bonds betwee complementary base pairs, and split the DNA into 3 polynucleotde strands- denaturing

Question 2

What do we drop the temperature to?

Answer 2

= drop to 55 degrees
= so that primers can attach- short sequence of single stranded DNA complementary to start and end of the dna fragment sequence (annealing)- cool enough for hydrogen bonds to reform]
= enzyme DNA polymerase aracges complementary free nucleotides and makes a new strand next to each template, phsoohodiester bond
= 2 copies of dna fragment so doubles in size in each cycle
= temperature is increaed to 72 degrees for this stage optimum for dna polymerase

Question 3

What are the advanages?

Answer 3

= automated
= rapid
= doesnt require living cells

Question 4

What is the equation used for pcr?

Answer 4

= numger of dna fragments produced
2^x