Dna

Question 1

DNAase/RNAase

Answer 1

free water acts as the diluent.

Question 2

Buffer

Answer 2

Acts to maintain an optimal pH environment

Question 3

Magnesium

Answer 3

Usually in the buffer, acts as a cofactor for polymerase activity

Question 4

dNTPs

Answer 4

nucleotide building blocks (A, T, G,

and C) to create PCR product.

Question 5

Forward and Reverse Primers

Answer 5

Targets area to be amplified and provides site of attachment for new dNTPs

Question 6

Taq Polymerase

Answer 6

Act as the enzyme to synthesize PCR product. It incorporates nucleotides to the new strand of DNA based on the laws of complementary base pairing, using the sample as the template.

Question 7

Nucleic Acid

Answer 7

Sample for template

Question 8

Amount of each substance in master mix

Answer 8

DNAase/RNAase -55ul
10 x PCR Buffer with Mg2+ -10ul
dNTP (1.25mM of each dNTP) -16ul
Forward/Reverse Primers 10ul
Taq Polymerase- 1ul
Nucleic acid template- 8ul
 100ul total

Question 9

Thermocycler

Answer 9

an instrument that will cycle through temperature to create amplicons

Question 10

Denature

Answer 10

Approximately 90-96

Double stranded DNA becomes single stranded as hydrogen bonds are broken by heat

Question 11

Anneal

Answer 11

Approximately 50-70

Forward and reverse primers attach to template DNA around the target area to be amplified

Question 12

Extend

Answer 12

Approximately 68-75
This temperature allows for optimal activity of the Taq polymerase enzyme to synthesis PCR product.

Taq polymerase synthesized a new strand of DNA complementary to the template strand