Hi,
I am interested in creating cDNA libraries from soil rhizosphere samples, but I (and my lab) have no experience with this process. I have found products for generating ds-cDNA from RNA (looking at using the SuperScript ds-cDNA Synthesis Kit from Life Technologies, unless anyone knows of a better/cheaper one), but I am trying to figure out what I can do before this to reduce the tRNA and rRNA in my samples. I do not want to enrich specifically for eukaryotes (plants, fungi, or otherwise) or prokaryotes because I want to look at mRNA being produced from everything.
I found a Terminator 5' phosphate dependent exonuclease from Epicentre that will target the 16S, 18S, 23S, and 28S rRNAs, but it will not remove tRNA or 5S rRNA, which seem to be very abundant. I am sure this must be something people need to do routinely, and I was wondering if anyone had any advice for what to do?
Thank you in advance!
I am interested in creating cDNA libraries from soil rhizosphere samples, but I (and my lab) have no experience with this process. I have found products for generating ds-cDNA from RNA (looking at using the SuperScript ds-cDNA Synthesis Kit from Life Technologies, unless anyone knows of a better/cheaper one), but I am trying to figure out what I can do before this to reduce the tRNA and rRNA in my samples. I do not want to enrich specifically for eukaryotes (plants, fungi, or otherwise) or prokaryotes because I want to look at mRNA being produced from everything.
I found a Terminator 5' phosphate dependent exonuclease from Epicentre that will target the 16S, 18S, 23S, and 28S rRNAs, but it will not remove tRNA or 5S rRNA, which seem to be very abundant. I am sure this must be something people need to do routinely, and I was wondering if anyone had any advice for what to do?
Thank you in advance!
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