Hello All My Microbio Experts,
I'm a total noobie in this field so go easy...
I've extracted RNA from cells and the A280/A260 values look good (~2.0 for most samples) which means the RNA is pure. However, the A260/A230 values are very low (0.1 - 1) meaning the sample is contaminated (probably GuHCl??). What can I do to retain the sample and get that A260/A230 value up before I do RT and then finally qPCR? Is this what ethanol precipitation does? If so does anyone have a procedure they could share?
Thanks!
Mike
Hi,
Looks like you may have phenol contamination in your extracted RNA. Re-purifying the RNA should help you get rid of the contamination while not compromising the purity. You can use either chloroform-isopropanol precipitation or Qiagen's RNA clean up kit.
Here is a protocol which might help for chloroform/isopropanol precipitation method-
Article Purification of Nucleic Acids by Extraction with Phenol:Chloroform
Hope this helps!
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