Hello,
To check specificity of your primers by ncbi blast, we find identity percentage, query cover and E-value as the parameter to be taken care off. Which parameter is the most important and given careful attention?
Identity percentage and Query coverage are very important because they both are defining your sequence specificity.
For primer designing query cover 100% would be better.
I'd use the design tool for this, which automates this for you, and tells you more directly what you care about. http://www.ncbi.nlm.nih.gov/tools/primer-blast/
Identity percentage and Query coverage are very important because they both are defining your sequence specificity.
For primer designing query cover 100% would be better.
This factors are imoprtant.
You must design a primer by primer 3 or generunner or pickprimer in ncbi using bacteria genome and then recheck primers with primer-blast in pubmed
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