MP-Methy BETA : Primer Design for Bisulfite Sequencing and Methylation-Specific PCR

Introduction Bisulfite Sequencing Methylation-Specific PCR

Target settings

Primers for bisulfite sequencing will avoid CG dinucleotides, while those for methylation-specific PCR will require a minimum number of CG sites in the 3' end region.
bp
Length (from the primer's 3' end) where CG dinucleotides are not allowed.
Max
Maximum number of CG dinucleotides allowed in the primer sequence.

Quality control settings

Min
°C
The minimum binding stability between primer and its binding sites.
From
To
Product size: 0 (min) - 1000000 (max).
Min
Max
Number of non-specific amplicons.
Min
kcal/mol
Primers with dimer stability exceeding this threshold will be discarded.
Min
Primers with hairpin stability exceeding this threshold will be discarded.
Min
Minimum number of different bases between target amplicon and off-targets if allowed.

Primer settings

bp
bp
bp
%
%
%
°C
°C
°C

Product settings

%
%
%
bp
bp

Thermodynamic settings

mM
mM
mM
nM

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