Ratio of 260/280 ~ 1.80, is really much matter for PCR or RT PCR ? Or ~2.00 will also works for the same.
A 260/280 ratio of 1.8 is considered to be pure enough for most practical purposes. While 2.0 is in theory even better it shouldn't significantly effect your PCR.
A 260/280 ratio of 1.8 is considered to be pure enough for most practical purposes. While 2.0 is in theory even better it shouldn't significantly effect your PCR.
@ Muhammed Mallah .. Sir DNA Isolation from Sputum samples for Mycobacterium Tuberculosis.
The 260/280 ratio is totally irrelevant for analysing purity of a nucleic acid preparation. If you check, you will discover that this ratio was designed to detect nucleic acid contamination of protein preparations, for which is it extremely sensitive. Protein contamination of DNA, on the other hand, is not and a ratio of 1.8 can result from up to 40% contamination with protein.
For (RT)-PCR all that matters is that your sample does not inhibit either the RT or the PCR and you can check this most easily by comparing Cqs from a 10-fold dilution: if the Cqs differ by less that 3.3, you are seeing inhibition.
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