Examine the diagram of a PCR reaction below and then answer

Examine the diagram of a PCR reaction below, and then answer the questions that follow. You may work in groups. After all groups have answered the questions, each group will take turns explaining the answers for the questions in front of the entire points for participation and questions. Students should keep the laboratory with correct answers since several questions on Exam 3 will come from this lab exercise. Taq polymerase 10X buffer 4 deoxy nucleotides dATP, dGTP, dCTP, dTTP two each of HL pNmers DNA template 2HL. i HL each 8 HL PCR reaction tube Sterile dl water 20HL total volume

Solution

It the target DNA template number is 2 then the formula for the number of copies of DNA obtained after \'n\' cycles = 2⁽ⁿ⁺¹⁾

In the question the target number is 100, so

after 1 cycle the number of target copies =100⁽¹⁺¹⁾ = 10000 copies of target

2) After 5 cycles the number of target copies = 100⁽⁵⁺¹⁾

= (100)⁶ = 10¹² copies

4) After 20 cycles-

100⁽²⁰⁺¹⁾ = 10⁴² copies

After 30 cycles it is 10⁶²

5)

5’ TTGACCATGTACATTACAGAATCCCATGCTAGCGGATCGATGCTAGCATT 3’

3’AACTGGTACATGTAATGTCTTAGGGTACGATCGCCTAGCTACGATCGTAA 5’

Primers

5’ TTGACCATGTACATTACAGAATCCCATGCTAGCGGATCGATGCTAGCATT 3’

                                                                       3’ATCGCCTAGCTACGATCG5’

        5’ACCATGTACATTACAGAAT 3’                                                   3’AACTGGTACATGTAATGTCTTAGGGTACGATCGCCTAGCTACGATCGTAA 5’

After 1^st cycle

5’ACCATGTACATTACAGAATCCCATGCTAGCGGATCGATGCTAGC3’

3’TGGTACATGTAATGTCTTAGGGTACGATCGCCTAGCTACGATCG5’

5’ACCATGTACATTACAGAATCCCATGCTAGCGGATCGATGCTAGC3’

3’TGGTACATGTAATGTCTTAGGGTACGATCGCCTAGCTACGATCG5’

5c) if there are no primers no amplification will occur.

5d) If G nucleotide is not added, then the PCR product will not be formed