Primer walking
This sequencing method is ideal for:
Principle
The use of several primers makes it possible to obtain overlapping sequences leading to the reconstitution of the complete product. A universal primer is used to sequence an insert, obtaining a fragment of the vector sequence and the beginning of the insert.
A specific primer can then be designed on the sequence obtained and used to obtain the rest of the sequence. If the sequence of interest is known, all the sequencing reactions can be carried out simultaneously. This service can be performed either in single or double reading.
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Mutation detection
Sanger sequencing can be adopted to verify
Principle
Among other applications, the mutation detection is used to monitor TILLING (Targeting Induced Local Lesions in Genomes) in plants, or to verify mutations in genes of interest.
We can design specific primers to target a mutation and compare it to a reference sequence provided for verification.
If several mutations/positions are to be verified in the same genome region, we can also produce a detailed variance report compared with a reference gene.