Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques > Agarose Gel Electrophoresis Forum
Register Search Today's Posts Mark Forums Read

Agarose Gel Electrophoresis Forum Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


Dnase contamination in plasmid mini prep

Dnase contamination in plasmid mini prep - Agarose Gel Electrophoresis Forum

Dnase contamination in plasmid mini prep - Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 08-19-2010, 06:56 PM
Pipette Filler
Points: 215, Level: 4 Points: 215, Level: 4 Points: 215, Level: 4
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2010
Posts: 4
Thanks: 0
Thanked 1 Time in 1 Post
Question Dnase contamination in plasmid mini prep



I have been having a problem where I have isolated DNA from dna prep, nanodropped the sample to confirm actual presence of DNA and absence of contaminants, then ran on a gel and had really huge blotchy bands show up far under my ladder. I believe i have figured out that I have dnase contamination which is chopping up my plasmid. I was wondering if anyone had an idea of the source of dnase contamination???????
Method:
all glass & plastic was sterile
ligation was good
plasmid was amp/carb resistant and colonies were selected
colonies were grown overnight w/ antibiotic
miniprep was done (all reagents were new and have worked with other lab workers preps)
dna was ran on a 1% gel, plasmid expected to run around 9kb
gell ran showing ladder showing control vector plasmid but sample plasmid band ran below the 1kb ladder under 500bp

I am going to try a phenol/chloro extraction. Again, I am just wondering if anyone has any idea about how dnase contaminated my sample!?
Reply With Quote
  #2  
Old 09-02-2010, 04:22 PM
Pipette Filler
Points: 105, Level: 2 Points: 105, Level: 2 Points: 105, Level: 2
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2010
Location: kurdistan
Posts: 5
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Dnase contamination in plasmid mini prep

try to load ur DNA again with appropriate loading dye
Reply With Quote
  #3  
Old 05-13-2011, 07:45 PM
Pipette Filler
Points: 2,551, Level: 32 Points: 2,551, Level: 32 Points: 2,551, Level: 32
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2009
Posts: 21
Thanks: 0
Thanked 3 Times in 3 Posts
Default Re: Dnase contamination in plasmid mini prep

that's just ccc conformation of your plasmid
Reply With Quote
Reply

Tags
contamination , dnase , gel electrophoresis , mini , plasmid , prep


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
low LPS plasmid prep protocol Bari Zahedi Protocols and Methods Forum 12 08-26-2009 01:37 AM
Poor Plasmid Prep Yield zek Molecular Cloning Forum 0 02-10-2009 07:45 PM
pGEX 2T vector maxi prep or mini prep david1259 DNA Techniques 2 11-02-2008 01:48 PM
Plasmid prep issues peter Protocols and Methods Forum 4 04-07-2007 10:53 PM
DNase digest during protein prep tin_tin Protocols and Methods Forum 1 11-03-2004 03:17 PM


All times are GMT. The time now is 10:07 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12280 seconds with 16 queries