QPCRprimer version 1.2 (2002-2006)
written by Peter Bagossi
Find QPCR primers in a given DNA sequence:
Salt concentration [M]:
Sequence:
Important notes:
Maximum number of characters in one line: 250
Maximum number of input lines: 1000
Maximum number of nucleotides: 500000
Limits based on "Alkami Biosystems Primer Design Tips" or "Sambrook: Molecular cloning"
Equation used:
Tm=81.5+16.6log(salt/(1+0.7salt))+0.41(%GC)-500/N
length of primers: min: 15 max: 30
%GC content: min: 35 max: 65
Tm of outside primers: min: 57.0 max: 60.0
Tm of inside primer: min: 67.0 max: 70.0
Distance between outside primers: min: 15 max: 90
Distance between outside and inside primers: min: 1 max: 5
Outside primers will start or end with A or G
Inside primer will start or end with A
Created: March 22, 2006
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Updated: March 28, 2006