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Problem 4 In the gene shown below (5'-3'), four possible primer sites are shown in bold for PCR amplification of the target DNA sequence hilighted in green. Three considerations for choice of primer are: (a) primer length; (b) GC content and Tm; and (c) lack of undesired complementarity. Here are two companies' websites describing tips for choosing PCR primer sequences: https://www.neb.com/en-us/nebinspired-blog/proven-tips-for-pcr-primer-design https://www.thermofisher.com/blog/behindthebench/pcr-primer-design-tips/ TTAATGTGAG TTAGCTCACT CATTAGGCAC CCCAGGCTTT ACACTTTATG CTTCCGGCTC GTATGTTGTG TGGAATTGTG AGCGGATCAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGGATT CACTGGCCGT CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGATC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGCG CTTTGCCTGG TTTCCGGCAC CAGAAGCGGT GCCGGAAAGC TGGCTGGAGT GCGATCTTCC TGAGGCCGAT ACTGTCGTCG TCCCGTCAAA CTGGCAGATG CACGGTTACG ATGCGCCCAT CTACACCAAC GTGACCTATC CCATTACGGT CAATCCGCCG TTTGTTCCCA CGGAGAATCC GACGGGTTGT TACTCGCTCA CATTTAATGT TGATGAAAGC TGGCTACAGG AAGGCCAGAC GCGAATTATT TTTGATGGCG (a) From the four bolded sites, choose two sites for PCR primer design and indicate why the other choices are would likely lead to an unsuccessful result. (b) Draw your two primer sequences (in the 5'- to 3'- direction) that you would purchase to amplify this target sequence by PCR.

Problem 4 In the gene shown below (5'-3'), four possible primer sites are shown in bold for PCR amplification of the target DNA sequence hilighted in green. Three considerations for choice of primer are: (a) primer length; (b) GC content and Tm; and (c) lack of undesired complementarity. Here are two companies' websites describing tips for choosing PCR primer sequences: https://www.neb.com/en-us/nebinspired-blog/proven-tips-for-pcr-primer-design https://www.thermofisher.com/blog/behindthebench/pcr-primer-design-tips/ TTAATGTGAG TTAGCTCACT CATTAGGCAC CCCAGGCTTT ACACTTTATG CTTCCGGCTC GTATGTTGTG TGGAATTGTG AGCGGATCAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGGATT CACTGGCCGT CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGATC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGCG CTTTGCCTGG TTTCCGGCAC CAGAAGCGGT GCCGGAAAGC TGGCTGGAGT GCGATCTTCC TGAGGCCGAT ACTGTCGTCG TCCCGTCAAA CTGGCAGATG CACGGTTACG ATGCGCCCAT CTACACCAAC GTGACCTATC CCATTACGGT CAATCCGCCG TTTGTTCCCA CGGAGAATCC GACGGGTTGT TACTCGCTCA CATTTAATGT TGATGAAAGC TGGCTACAGG AAGGCCAGAC GCGAATTATT TTTGATGGCG (a) From the four bolded sites, choose two sites for PCR primer design and indicate why the other choices are would likely lead to an unsuccessful result. (b) Draw your two primer sequences (in the 5'- to 3'- direction) that you would purchase to amplify this target sequence by PCR.

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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help me with part a and b please

Problem 4 In the gene shown below (5'-3'), four possible primer sites are shown in bold for PCR amplification of the target DNA sequence hilighted in green. Three considerations for choice of primer are: (a) primer length; (b) GC content and Tm; and (c) lack of undesired complementarity. Here are two companies' websites describing tips for choosing PCR primer sequences: https://www.neb.com/en-us/nebinspired-blog/proven-tips-for-pcr-primer-design https://www.thermofisher.com/blog/behindthebench/pcr-primer-design-tips/ TTAATGTGAG TTAGCTCACT CATTAGGCAC CCCAGGCTTT ACACTTTATG CTTCCGGCTC GTATGTTGTG TGGAATTGTG AGCGGATCAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGGATT CACTGGCCGT CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGATC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGCG CTTTGCCTGG TTTCCGGCAC CAGAAGCGGT GCCGGAAAGC TGGCTGGAGT GCGATCTTCC TGAGGCCGAT ACTGTCGTCG TCCCGTCAAA CTGGCAGATG CACGGTTACG ATGCGCCCAT CTACACCAAC GTGACCTATC CCATTACGGT CAATCCGCCG TTTGTTCCCA CGGAGAATCC GACGGGTTGT TACTCGCTCA CATTTAATGT TGATGAAAGC TGGCTACAGG AAGGCCAGAC GCGAATTATT TTTGATGGCG (a) From the four bolded sites, choose two sites for PCR primer design and indicate why the other choices are would likely lead to an unsuccessful result. (b) Draw your two primer sequences (in the 5'- to 3'- direction) that you would purchase to amplify this target sequence by PCR.
Transcribed Image Text:Problem 4 In the gene shown below (5'-3'), four possible primer sites are shown in bold for PCR amplification of the target DNA sequence hilighted in green. Three considerations for choice of primer are: (a) primer length; (b) GC content and Tm; and (c) lack of undesired complementarity. Here are two companies' websites describing tips for choosing PCR primer sequences: https://www.neb.com/en-us/nebinspired-blog/proven-tips-for-pcr-primer-design https://www.thermofisher.com/blog/behindthebench/pcr-primer-design-tips/ TTAATGTGAG TTAGCTCACT CATTAGGCAC CCCAGGCTTT ACACTTTATG CTTCCGGCTC GTATGTTGTG TGGAATTGTG AGCGGATCAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGGATT CACTGGCCGT CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGATC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGCG CTTTGCCTGG TTTCCGGCAC CAGAAGCGGT GCCGGAAAGC TGGCTGGAGT GCGATCTTCC TGAGGCCGAT ACTGTCGTCG TCCCGTCAAA CTGGCAGATG CACGGTTACG ATGCGCCCAT CTACACCAAC GTGACCTATC CCATTACGGT CAATCCGCCG TTTGTTCCCA CGGAGAATCC GACGGGTTGT TACTCGCTCA CATTTAATGT TGATGAAAGC TGGCTACAGG AAGGCCAGAC GCGAATTATT TTTGATGGCG (a) From the four bolded sites, choose two sites for PCR primer design and indicate why the other choices are would likely lead to an unsuccessful result. (b) Draw your two primer sequences (in the 5'- to 3'- direction) that you would purchase to amplify this target sequence by PCR.
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