PCR Primer Designing | NCBI Primer BLAST | In silico PCR primer designing and validation 

In this video we will design a primer using NCBI Primer BLAST
PCR is a commonly used method to amplify DNA of interest in many fields such as biomedical research, diagnostic testing and forensic testing. While the outcome of PCR can be influenced by many other conditions such as the template DNA preparation and reaction conditions, designing a good pair of primers is a critical factor, A general requirement is that primers should have similar melting temperatures (Tm) and a balanced G/C content, but should avoid self-complementarity and hair-pin structure.
A number of public software tools have been developed to aid the primer design process. Notably, the widely used Primer3 program designs primers based on a variety of parameters. Since it does not perform target analysis, users typically need to test the primer specificity using additional tools. However, this process is time-consuming and sometimes even impractical if the primers have too many database matches. NCBI Primer BLAST is vergood tool, it not only gives us a predicted primer list, but also provides a specific target analysis.
NCBI Primer BLAST: www.ncbi.nlm.nih.gov/tools/pr...
Primer Stat: sites.ualberta.ca/~stothard/j...
Reverse Complement: www.bioinformatics.org/sms/re...
Literature": link.springer.com/chapter/10....
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