Bacterial mRNA from environmental samples - (May/11/2006 )
I've successfully extracted total RNA from environmental samples (as demonstrated by clear bands on the gel and high A260/A280), but I could not get any product from RT-PCR. I wonder if there is any good suggestion for a good kit on bacterial mRNA extraction.
Any suggestion is much appreciated.
I Remember reading a few paper on the subject (I was interested in extracting mRNA from manure). The conclusion was that no kit is really good and that you should use different extraction methods successively. The papers are at the lab so I can't give any reference now, but I will look that up tomorrow.
A kit may work in your case to purify the fraction of smaller RNA(mRNAs). Most kits ability to extract RNA may vary between kits and sometimes between samples. So, your cDNA conversions may not be as accurate or robust. Look for a kit that is specifically designed to purify small RNAs. What kits have you tried?
In the lab now, here is what I had found, in AEM. Hope this helps.
Thanks a lot for the paper and suggestions.
I've tried to extract total RNA using TRIzol and purified with RNeasy column. Then I used Quantitect of Qiagen to do RT-PCR. Unfortunately, I could not get clear bands, all I got was smearing bands.
Any thoughts on this? I am thinking about checking the Invitrogen Superscript III First strand synthesis kit, has anyone checked this?