Mixed Culture 16s rRNA


by HOMO
Tags: 16s ribosomal rna, 16s rrna
HOMO
HOMO is offline
#1
Nov1-13, 11:45 PM
P: 4
I have a mixed culture plate and I want to do 16s rRNA analysis on it and I have a few questions.

First of all there are no visible colonies, but I know bacteria is growing. Do I just take a small chunk of the medium and do PCR on it with universal primers (I have a procedure for this step)?

Second, because it is mixed culture, wont the PCR be an amplification of a bunch of different 16s from the different bacteria? How do I separate the strains? I have an idea how to do this. After PCR on the universal primers, I can run it out on a gel and excise the separate fragments, then purify so that I have pure samples of all the amplified RNA?

Third, after I purify the RNA, I send it to get sequenced and then I use some database such as BLAST to see what I have?
Phys.Org News Partner Biology news on Phys.org
Citizen scientists match research tool when counting sharks
Microbes provide insights into evolution of human language
First sex determining genes appeared in mammals 180 million years ago

Register to reply

Related Discussions
16S rRNA sequence information to identify organisms Biology 1
07/28/08 PHD comic: 'Pop Culture!' Science Comics 1
Albertros culture Social Sciences 5
Could the culture war become civil war? Current Events 98
our culture, Gen X General Discussion 3