One of the uses: to compare the PCR product to your original template to see whether any base mutation occur after PCR reaction. The mutation rate will be different depending on which kind of Taq DNA polymerase you use. Less mutation occurs when High Fidelity Taq DNA polymerase is used.
One of the uses: to compare the PCR product to your original template to see whether any base mutation occur after PCR reaction. The mutation rate will be different depending on which kind of Taq DNA polymerase you use. Less mutation occurs when High Fidelity Taq DNA polymerase is used.
As already answered by Mr. Yau, sequencing can be done for checking mutations or simply polymorphisms in the PCR-amplified sequence.
In addition to all that has been said, PCR can be used to perform screening after cloning (detection of recombinant clone ) and sequencing of the target DNA (insert) can be done to identify for example strains.
You can use the result of sequencing for genotyping and phylogenetic analysis too.
There are many uses for PCR reactions. As mentioned, one goal may be phylogenetic approach : amplifying all the sequences located between the two primers that have been chosen (corresponding to the conserved parts of an evolutionary conserved gene with some polymorphism located between the conserved primers). This is typically what is done with ribotyping. Sequencing the PCR products allows one to defined the population of sequences and thus organisms that are present in the extract, with possibly novel species that may be defined depending on the score of alignment with sequences of already known and characterised organisms.
Another use: when you want to modify an allele (such as to make a null allele) in vivo by using Designer Nucleases (such as ZFN, TALEN,...), which creases double-stranded breaks in the gene-of-interest, you may want to check the final experimental outcome of the specific gene sequence to see whether or not the gene has been disrupted as planed through deletion or addition of bases. Then, PCR of the gene will be conducted and then sequenced.
Sequencing gives an idea of polymorphism or single nucleotide variants induce by DNA polymerase during PCR reaction.
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