Forward Primer : 5\'- GTGTAT AAGCTT _____________________________-3\' Reverse Pr

Question

Forward Primer : 5'-GTGTATAAGCTT_____________________________-3'
Reverse Primer: 5'-ATTTATGGTACC______________________________-3'

b). What will be the theoretical length of the expected PCR product to be amplified using your section primers? Specify the number of nucleotides

ORIGIN 1 ctgtaaaaca attttttaga tttgtaagca ctcaatttga atgcttttag tcttagttat 61 tttatttttc gtacggtaga tgggcgttta gtattttaac cgcatcacgg tcacactggc 121 ggatttgttt ttattttggt cgccatgcaa agtaaacacc gcaaactgct gctgcgctgc 181 ctcctggtgc tgcccctcat ccttctggtg gactattgcg gcctgctgac ccacctgcac 241 gagctgaact tcgagcggca ctttcactac ccgctgaacg atgacactgg atctgggtcc 301 gcatcctctg gactggacaa gttcgcctac ctgcgagtgc cctcgttcac ggcagaggtg 361 ccggttgacc agcctgcgcg gctcacgatg ctcataaaga gcgcggtggg caattcgcga 421 cgtcgggagg ccattcgccg gacttggggc tacgagggtc gattctcaga cgtccatctg 481 cgtcgcgtgt tcctgctggg cacggccgag gatagtgaga aagacgtggc ttgggagtcc 541 cgcgagcatg gcgacattct gcaggcggag ttcacggacg cctacttcaa caacacgctg 601 aaaaccatgc tgggtatgcg ctgggctagc gatcagttca atcgcagcga gttctatctt 661 ttcgtggacg acgactacta tgtttcggcc aagaacgtgc tcaagtttct gggcaggggg 721 cgtcagagcc accagccgga gttgctgttc gccgggcacg tgtttcagac atccccgctc 781 cgccacaagt tcagcaagtg gtacgtttcg ttggaggaat accccttcga tcggtggcca 841 ccgtacgtga ccgccggcgc gttcattctg tcgcagaagg cactgcgcca gctgtatgcc 901 gccagcgttc acctgccgct cttccggttc gacgacgtct acctgggcat agtggccttg 961 aaagcgggca tttcccttca gcactgcgac gacttccgct tccatcggcc ggcgtacaaa 1021 ggtccggaca gctatagttc ggtgatagcc tcgcacgaat tcggcgatcc cgaggagatg 1081 acgcgggtat ggaacgagtg ccgatccgcc aattacgcgt agcagcaaga acaggcagga 1141 ctgaagagcg ctgcagattg tacatatata cgaacgagac agcacctaga caacagcctt 1201 ataaagtata ttaggagcag tggatgatac ctcaaccgtt cagtgagact ttaaaccaag 1261 cagctggtgc taatagtgca agatcgcaag agtaattcaa ttctaattgt aagcgtaatg 1321 ataaagcagt tgattcaatt cgtatatata gagattatgt gcaaatgtcg gataacgtat 1381 gtgtggtctg atacctttga atgcaatata aagttgatta acataggaaa gcgaataaaa 1441 aacttaaaat

Explanation / Answer

1.To design a reverse primer

5' TAA AAT TCA AAA AAT AAG CG 3'

5' ATT TTA AGT TTT TTA TTC GC 3'

5'-ATTTATGGTACC_ATT TTA AGT TTT TTA TTC GC_____________________________-3'

2. To design a forward primer

5’ CTG TAA AAC AAT TTT TTA GA3’

5'-GTGTATAAGCTT_CTG TAA AAC AAT TTT TTA GA____________________________-3'

3 Length of PCR product.

The PCR product that is the amplicon will contain both the primer regions and the gene sequence in between them. So, it will be 1441 nucleotides of given DNA sequence as can be seen in the left bottom corner , plus 12 nucleotides each of the designed forward and reverse primer sections given in the question (Forward Primer: 5’ GTGTATAAGCTT 3’ and Reverse Primer: 5’ ATTTATGGTACC 3’). The total becomes 1465 nucleotides.

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