In article <jsg-1907961315480001 at lacto.path.cam.ac.uk>, jsg at mole.bio.cam.ac.uk (James Good) says:
>>Is it possible to make cDNA libraries from gram negative bacteria? Some
>genes have short poly-a termini, so perhaps it would be possible in the
>conventional way, but is there a more reliable method? Thanks.
The m-RNA in eubacteria do not have the poly A tails that are found in
eukaryotic cells. The primary reason to make cDNA libraries from
mammalian cells is to obtain DNA in which the introns have been removed.
This is not an issue with eubacteria. It is possible that there could
be situations in which it is desired to make a differential expression
library, e.g., same bacteria grown in two different environments.
Isolating m-RNA from bacteria for any reason is a difficult procedure,
as the half life is very short (minutes) and the material is rapidly
degraded. Procedures do exist to minimize degradation of m-RNA and are
used primarly for Northerns; so if one really had to make an expression
library, there are ways to do this.
Tom Dougherty
