i did RNA extraction using differnt protocol triazol and invetrogen RNA mini kit .. the 260/280 ratio was 1.9 to 2.1 but for the 260/230 i got 1.5 to 1.8 in both protocols, should i consider it for RT-PCR.. ??
The short answer is Yes! Go ahead, you will get results. Just make sure you include a control primer pair for some housekeeping gene(s) like actin or gapdh which will allow you to normalize and account for any small differences in in RT or PCR efficiency.
It is heavily dependent on the concentrations. It is noted that concentrations of guanidine thiocyanate of up to 100 mM in an RNA sample do not compromise the reliability of downstream applications.
(https://www.qiagen.com/us/resources/faq?id=c59936fb-4f1e-4191-9c16-ff083cb24574&lang=en)
Also check others opinion in previous raised questions.
https://www.researchgate.net/post/How_can_I_improve_RNA_quality_with_low_260_230_ratio
Thank you all for the helpfull answer ..
Hello fawaz.
what did u mean by adding dd-h2o .. which step?
i usualy normlaiz my sample concentration at the cDNA synthesis step to be ( 50ng/ul in 10 ul) to have 500 ng in 10 ul
dear Abhishek Kumar do you mean the sample conc. ?
it is all between 356 to 500
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