Question

A number of different buffers/components are used in the PCR. Explain the function of each: a. DNA plasmid b. Primer pair C. dNTPs mixture d. Taq polymerase

Answer 1

Q.1 A. Dna plasmid have no free ends, the are naked , with circular ends, it is naked double standard dna. It have RNA polymerase enzyme and proteins.

Function:- 1.they replicate independently,

2. they are origin of replication,

3.they provide sexual conjucation between cells,

4.they have trans gene- which transfer from one bacterium to other.

B. Primer pair :-  

function:-

1. They demarcate the segment .

2. They amplified dna templates .

3. They matched dna segments .

C - DNTPS MIXTURE :- It is basically sodium salt of solutions datp ,dctp , dgtp ,dttp.

FUNCTION:-

1. WITH the coordination with taq dna polymerase it expand the growth of dna stand .

2. With the help of hydrogen bond it get attached to its complementary dna strand .

3. it is used as bricks .

4. If DNTPS are in lower concentrations then their is increase in specificity of reaction .

D :- TAQ POLYMERASE

FUNCTION-- it helps in production of multiple copies by amplifying dna .

2. It is used in higher temperature also it do not loss it's significance.

3. It denature the dna duplex .

Q.2. the annealing temperature --

Basically it's should be in between 48 to72 degree celcius.

Some time many use annealing temperature must be below 5 degree Celsius.

It should be calculated as melting temperature it begins at 3 to 5 degree Celsius.