Found in some articles that Carrier RNA enhanced the yield of DNA from a variety of sources, so I tried for the first time during my DNA extraction from tiger faecal samples, but some unusual results appeared in my mitochondrial PCR. I am using Qiagen Stool Mini Kit and got the lyophilized carrier RNA from Qiagen as well. If anyone put few lines on the dilution of the carrier RNA (whether RNase free water or buffer from Qiagen?) and at which step to add the same during extraction process -- would be great.
Regards, Aziz
usually you use rna as a co-precipitant . When there is not much dna there may not be enough to form a precipitate to spin down after adding salts and alcohol and it just gives a large enough residue to trap the dna and allow it to stick to the bottom of the tube after centrifugation so I would add at the end of the extraction in case it interfered with the purification kit and stopped the dna binding. .If you think that the rna interferes with your PCR or then rnase treatment before the pcr would eliminate this effect. Or you could test the original rna to see if you get the same effect on your pcr
I distant times, we used glycogen as a co-precipitant when we had tiny amounts of DNA.
I don't think that would interfere with PCR, but might be wrong!
Geoff is right glycogen is a very good co-precipitant and I am sure that it does not interfere with pcr and very unlikely to be contaminated with DNA or to interfere with the measurement of amounts of DNA at 260nm wavelength
Hi Paul and Geoff, great! Many thanks for your kind discussion on the issues, hopefully be useful to come out the issues around carrier RNA.
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