Dear Researchers,
I have extracted the RNA by using the Qiangen Kit and also Tiangen Kit , by using passion fruit Peel at ripening stages, But the concentration is about 20 -50 ng/ul . A260/A280 1.98- 2.01,
I have tried my best to get higher concentration but failed, so my question is that
Low concentration with high quality RNA can use for cDNA and qPCR?
as because in cDNA protocl they mentioned that can take only 7ul maximum RNA, while other 3ul reagents, so in 20ng/ul RNA concentration how to convert to cDNA?
Md Abdur Rahim but when i willl have to prepare cDNA then use 1000ng or 500 ng or 300 ng or 200ng?
in cDNA kit the maximum RNA column can be use 7 ul , and other 3ul reagent . mean if i use 1000ng /200 RNA conc = 5ul RNA + 2ul Water,
but now in the current concentration 20-50 ng/ul what should i do for cDNA preparation>? you can have a look at the attached protocol of available kit
and if i will have 200 concentration RNA, and 20 ng/ul concentration RNA then there qPCR can run together ? or need to prepare separate cDNA/qPCR samples for 20 and separate for 200ng/ul?
if together then how can make same concentration of cDNA ?
Hi!
I agree with Md Abdur Rahim
It is possible. Try Quantitect reverse transcription kit from (Qiagen). It allows reverse transcription for loads0 of 10pg - 1ug of RNA per reaction.
Best,
Jacó
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