Login

Join for Free!
113932 members


First time making a genomic library

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderator: BioTeam

First time making a genomic library

Postby Awol » Mon Sep 17, 2012 10:41 am

I am a molecular biology student and I have been making genomic libraries in E.coli for the first time. I was posed a question by my prof. that has confused me.

I am asked whether this approach, using two different restriction enzymes, could be used to generate a complete genomic library? And if not why not. It is perhaps the definition of 'complete genomic library' that puzzles me, I would be grateful if someone could shine some light on this for me.

We are using the plasmid pUC19 with restriction enzymes EcoRI and BamHI.
Awol
Garter
Garter
 
Posts: 13
Joined: Fri Dec 03, 2010 2:20 am

Postby Cat » Mon Sep 17, 2012 1:23 pm

Possibly yes. But how would you know if it's complete? That I see as a problem.
Cat
King Cobra
King Cobra
 
Posts: 625
Joined: Thu Feb 14, 2008 7:40 pm

Postby JackBean » Tue Sep 18, 2012 10:33 am

You should get complete library (if you use sufficient number of clones) with any REs. There is a calculation to get number of clones which you have to screen to get your gene of interest with some probability.

The only thing which might be of concern could be telomeric ends of chromosomes and if these two REs cut too far appart. I'm not sure, what is the limit for the inserts, but it might happen, that they cut too far appart and thus such piece could not be used.
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
User avatar
JackBean
Inland Taipan
Inland Taipan
 
Posts: 5654
Joined: Mon Sep 14, 2009 7:12 pm



Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 2 guests

cron