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PCR primer design

PCR primer design - PCR - Polymerase Chain Reaction Forum

PCR primer design - 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 01-11-2012, 11:44 AM
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Default PCR primer design



My dears,

What is the tips for good designing of specific primer.
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Old 02-15-2012, 04:20 AM
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Default Re: PCR primer design

Hi there
please see this page:

http://www.molecularstation.com/pcr/pcr-primer-design/

Some tips on primer design

  • PCR primers are optimally designed as 15-30 oligo nucleotides in length
  • Primer oligonucleotide GC content should be between 40-60%
  • Primers should not be self-complementary or complementary to the other primer in the reaction mixture
  • melting temperature Tm of primers should not have a difference more than 5°C.
Use Webprimer or primer3 to design, then check Tm (melting temp), self-annealing and GC to make sure its ok.
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Old 01-16-2013, 05:44 AM
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Default Re: PCR primer design

[Only registered users see links. ] Enter one or both primer sequences in the Primer Parameters section of the form. If only one primer is available, a template sequence is also required. See "A Target Template Sequence.
In the Primer Pair Specificity Checking Parameters section, select the appropriate source Organism and the smallest Database that is likely to contain the target sequence. These settings give the most precise results. For broadest coverage, choose the nr database and do not specify an organism.
Click the "Get Primers" button to submit the search and retrieve template and specificity information.
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Old 02-22-2013, 04:48 PM
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Default Re: PCR primer design

just choose primer3 software. Itīs free and primers will work in about 80%.
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