5) Design a set of primers that will amplify the Gumby gene from Chromosome S. Y

Question

5) Design a set of primers that will amplify the Gumby gene from Chromosome S. Your primers should not be significantly complementary to each other, should be between 21-33 bp long. an have a melting temperature between 55 to 68 degrees 4 pts Calculate the melting temperature of the oligonucleotides with the following formula: Tm-(wA+xT) *2+ (yG+zC)*4 where w-# of As x-# of Ts etc Prediet the size of the resulting PCR product 5' 1 gacgcgegcg gaagcgctct gogggeccte ggaaaccgcc ceggcggctg agaggetgog 61 gccactgcct ggcaccecga cgggaggggc tocggatogt toggagcogg etgaacccct 121 toggecgcga gegacogeat gagtcggggg actatgeccc agccogaage gtggecaggc 81 gcgagetgcg cogagacgcc ggegegggag geggcggeca eggegeggga eggegggaag 241 geggcggcca gcgggcagcc geggeccgag atgcagtgcc cggccgagca tgaggaggac 301 atgtacegtg ctgcagatga aatagaaaag gagaaagaat tgcttataca tgaaagaggg 361 gcatcagaac cgagattaag cgtagetect gaaatggata tcatggacta ctgcaaaaaa 421 gaatggagag gaaatacaca gaaagcaacg tgtatgaaaa tgggetatga agaggtttet 481 cagaagttca cctccatacg gcgagtccgt ggtgataatt actgtgcact gagggccacg 541 ctgttccagg ccatgagcca ggetgtgggg etgecgccct ggctgcagga cccggagcte 601 atgctgttac cagaaaaact cataagcaaa tacaactgga teaagcaatg gaaacttgga 661 etgaaatttg atgggaagaa tgaggacctg gttgataaaa ttaaagagtc cettactctg 721 ctgaggaaga aggttgaaat gttecttctt gectatgetg tgegccacac catccaggtg 781 taccggctct ccaagtacaa cacggaagaa ttcatcacag totaccccac egacccaccc 841 aaggactggc cagtggtaac getcattgct gaggacgatc ggcactataa catecccgtc 901 agagtgtgtg aggagaccag tctatdagag acgcatgetc ctgacagect ggcgacgtgg 961 egaagatgca caggtggcte ctgggcttgg gctgcaggtt tgggggtetc taagaacaat 1021 ctctgagaag aacccttggg cccctgggag ccaagttgga caggatgtcc tgaagactag 1081 cttttgataa gagaaattaa ccaagtcttt ceectcatct atgatgcaat atatttcagt 1141 g9gggecttc agagcacacc tgttggacgg tgeaaaccat atcttctcca gaaggcaaat 1201 acttttgtat cagaggaaac tcagttttgg agaggaatat gttctttatg tetcaaatca 3 The coding sequence for the gene is believed to be 139 to 927

Explanation / Answer

SET OF PRIMERS :

forward primer : 5'catgagtcgggggactatccc3' (21bp )

its melting temperature (Tm)f =(wA+xT) *2 + (yG+zC)*4

WHERE, A=4 T= 4 G=7 C=6

Tm= 68 degree celcius.

reverse primer = 5'actagactggtctcctcacac 3' (21 bp long)

Tm = (wA +xT)*2 + "( yG+ zC)*4

where A=5 T=3 G=5 C=8

SO Tm = 64 degree celcius.

always remember primers are always written as 5' to 3' ( reverse and forward primer both )

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