I am new to PCR. I am extracting RNA from osteoblast cells and getting very low amount of rna. However i need to do pcr with 12 primers and i will be able to use 20 ng or less per reaction. Is it enough or do i have to look for other ways?
Hi,
It is OK.
you can find more information about the kit in the following link:
https://www.qiagen.com/us/shop/pcr/real-time-pcr-enzymes-and-kits/qpcr/quantinova-sybr-green-pcr-kit/#productdetails
The most important issue in RNA extraction is avoiding RNases, you must use RNase free water, tips etc. until you prepare the cDNA, after that it is stable.
Good luck!
Thank you Meisam,
I have gone through all the details. I have checked the purity of the RNA obtained and they were found to be of very good quality. I just wanted to confirm the if other people have done with less amount of rna. Thank you.
Cheers,
Urvashi
How much RNA ng has you obtained of what number of cells? You should do a pcr test using differents concentrations of RNA, closely to recomendated concentration for the kit protocol, for to know what is better management for your pcr. Maybe with less quantity of RNA you can do the pcr without problems, thus take up less cells.
Cheers.
Yes you can use. I have also use the concentration up to 10 ng in onestep RT and it work.
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