Poly (A) tail absent in bacterial m-RNA so oligo dT is not suitable for such purpose, random primer may be more suitable for this. . Can I use any kit have random primer? Are any kit available work perfectly for both eukaryotes and prokaryotes’ m-RNA for C-DNA synthesis?
Hi Ashraf
As far as I remember there are few kits/ or protocols available for selectively remove ribosomal RNAs and allows isolation of mRNA in bacteria....browse to see such products....If availavle after isolation of total mRNA you can even add a poly A tail using the enzyme poly A polymerase and then use cDNA synthesis primed with oligodT....A similar protocol is used for analysis of mature miRNAs..
check if it is available
bye
Hi Ashraf,
You can use Bioline cDNA synthesis Kit. it comes with random hexmers as well as oligo (dT) 18 primers. In fact most of the cDNA synthesis kits come with both primers.
good luck
Hi Ashraf. Independently of the kit that you decide to use, what I recommend is to use a reactive from Quiagen, RNAprotect Bacteria Reagent. In bacteria transcription, translation and almost mRNA degradation are happening simultaneously. This reactive provides immediate stabilization of RNA prior to RNA isolation procedure. It is specially critical with low abundant mRNA. In the past, I had some problems with poorly transcribed RNA and this is the only solution I found. After that, any normal kit with random hexmers should be Ok.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA