Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

shotgun library - small insert problem

shotgun library - small insert problem - Protocols and Methods Forum

shotgun library - small insert problem - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

LinkBack Thread Tools Display Modes
Old 02-16-2004, 05:48 AM
Krzysztof Lubieniecki
Posts: n/a
Default shotgun library - small insert problem

I am trying to construct shotgun library. To do this I isolate DNA
from BAC using standard alkaline lysis methodology. I shear DNA using
a sonicator. I always check results of sonication on the gel. If it's
ok, I run 1 % low melting gel and cut fragment between 2-5 kb. I
isolate DNA from the gel using Qiagen kit and do end repair reaction
using Epicenter end repair kit. I clean isolated DNA using Qiagen PCR
cleaning kit, do the ligation and after that transformation. I get
very good efficiency of transformation but my problem is that I get a
lot of small inserts, between 300 and 400 bp long (For 32 checked
plasmids I've got only 6 inserts above 1kb). Maybe someone had
similar problem and can suggest what can I do to decrease number of
small inserts.

Reply With Quote

insert , library , problem , shotgun , small

Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
qPCR NEWS - small activating RNA (saRNA) Editor www.Gene-Quantification.info Protocols and Methods Forum 0 02-26-2008 12:30 PM
DNA Cloning Protocol moleculardude Molecular Biology Articles and Protocols 2 08-05-2007 02:31 PM
Small problem... Green_Monkey23 Arabidopsis and Plant Biology 2 10-05-2006 06:08 PM
Simply put, MOVING DIMENSIONS THEORY is THE NEW MODEL: http://physicsmathforums.com drelliot@gmail.com Physics Forum 0 08-29-2006 06:32 AM
Sci.chem FAQ - Part 1 of 7 Bruce Hamilton Chemistry Forum 0 01-15-2004 09:06 AM

All times are GMT. The time now is 05:17 AM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2015, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12897 seconds with 16 queries