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Primer design for semi-random mutagenesis

Mutantprimers

Description

This tool designs QuickChange primers in bulk, either traditional or staggered, like the Deepscan tool, except that the amino acid mutations required are specified. This includes options for degerate codons (see below).

Input

To make primers at the ends of the coding sequence, the neighbouring noncoding sequence are required. Two options are available:

To give the upstream and downstream sequence separately.
To give a sequence with a start and end position. A variant of this is to give the starting/end position as a short sequence to match as opposed to a number.

Upstream
Sequence

Sequence

Downstream
Sequence

 
First base

Last base

Add to the following textarea the list of mutations desired, such as A13P. If more complex mutations, such as degenerate codons are to be use mark the destination codon in square brakets after the AA residue and its number, e.g. A13[NNK]. For mutations to multiple amino acids (e.g. A13GP) please specify the required codon for now. If considering degenerate codons and are not sure which to pick, try using the helper tool.



Mutation
List


Enzyme

Target Tm °C

GC clamp bonus °C

Method

Overlap nt

Length