Hello all. I would be grateful if I could use valuable comments from anybody who has experience in this field.
my nasopharynx swabs are not fresh. they have collected and immediately frozen at -70 untile December 2018 to May 2019 without using RNA later! the concentration of RNA which I read by nanodrop is around 45-50 and sometimes around 90-100 ng/mL .
I used RNase free Glycogen as a carrier to the aqueous phase before adding isopropanol but my concentration was around 53 ng/mL. Is there any way to improve the concentration of cellular RNA? and I want to know can I do syber green real-time pcr even on these low yield RNAs for a transcript quantification PCR?
Trizol I used: Invitrogen
Thank you
Looking forward to your valuable response as soon as possible.
Hello
If you ng/mL stands for nano-gram/milliliter then you are getting very very low RNA concentration, but if it means nano-gram/microliter then you are fine and you should not expect more concentration.
Regards
Ali Javadmanesh Hello
Thank you very much for your response. I mean nano-gram/microliter. I extracted RNA from another sample and I got 147nano-gram/microliter. So I think the cell concentration is very different in my samples.
Hello
Looks like it, the other possibility could be the efficiency of extraction. This is another variable to be considered. Although you do not seem to have a big problem regarding your concentration of extracted RNA.
Good luck
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA