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Which two primer pairs would work to PCR amplify the double stranded DNA target

Which two primer pairs would work to PCR amplify the double stranded DNA target - PCR - Polymerase Chain Reaction Forum

Which two primer pairs would work to PCR amplify the double stranded DNA target - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 07-15-2009, 12:19 AM
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Default Which two primer pairs would work to PCR amplify the double stranded DNA target



sequence shown below? Which two primer pairs would work to PCR amplify the double stranded DNA target sequence shown below
3' .. GGCTGAGTGCTGGATGAGTGGGCTGAGCTTGCTA...5'
5' ...CCGACTCACGACCTACTCACCCGACTCGAACGAT...3'

Would it be...
5' GGCTGAGT 3' and 5' TACGAAGC 3'?
5' TGAGTCGG 3' and 5' CGAACGAT 3'?
5' CCGACTCA 3' and 5' ATCGTTCG 3'?
or
5' TACTCACC 3' and 5' GGTGAGTA 3'?
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Old 07-16-2009, 12:51 PM
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Default Re: Which two primer pairs would work to PCR amplify the double stranded DNA target

First of all, I think you might need to read up a bit on the basic concept of PCR and how it works; it comes across as if you're new at this. Secondly, I would suggest you use a primer designing software. The one I use is PrimerPremier, and I find it pretty good.

It doesn't matter if your template is double-stranded, it will be denatured at 94 C. When denatured, your primers should (theoretically) bind to the single-stranded DNA and the elongation will ensue.

Therefore, to design primer pairs:
1. For template, typically we use the orientation of 5'->3'.
2. Find optimum primer pairs using the primer design software, which would show you the annealing temperature, and chances of false priming/hairpin formation.
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Old 07-17-2009, 08:54 AM
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Default Re: Which two primer pairs would work to PCR amplify the double stranded DNA target

example


5' ...ccgactcacg...3'
5' ...ccgactcacgacctactcacccgactcgaacgat...3'
________________________3'...tgagcttgcta...5'


thus to order for primer is
5' ...ccgactcacg...3' & 5'...atcgttcgagt...3'

Last edited by butters; 07-17-2009 at 08:57 AM.
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amplify , dna , double , pairs , pcr , primer , stranded , target , work


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