Owaiskhan9654/Gene-Sequence-Primer-
Gene and Primer Sequence Analysis for SARS-CoV-2, EGFR(Non Small Lung Cancer Cell), Influenza DNAs ### How can I check my Oligo primers to ensure there are no significant primer design issues? - The difference between melting temperatures (Tm) of the primers should be less than 5°C. - The GC content should be between 35-80% or equivalent to the product being amplified. - The Delta G value of any self-dimers, hairpins, and heterodimers should be weaker (more positive) than -9.0 kcal/mole. Positive numbers indicate that the actual secondary structure shown will not form at all. - Avoid 3' complementarity between the two primers to prevent primer dimers. The IDT OligoAnalyzer APIs can be used to assess these different criteria for a proposed oligo. #### [Reference](https://sg.idtdna.com/pages/support/faqs/how-can-i-check-my-pcr-primers-using-the-oligoanalyzer-program-to-ensure-there-are-no-significant-primer-design-issues-)
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