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

DNA Techniques Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


Why iam not able to get a band from my Plasmid DNA samples?

Why iam not able to get a band from my Plasmid DNA samples? - DNA Techniques

Why iam not able to get a band from my Plasmid DNA samples? - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 06-09-2009, 12:16 PM
Pipette Filler
Points: 7, Level: 1 Points: 7, Level: 1 Points: 7, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Why iam not able to get a band from my Plasmid DNA samples?



Hi friends
I had isolated plasmid DNA frm E coli and agrobacterium samples .. I had got more pellets after washing in absolute ethanol. But when i run the gel i was not able to see the DNA band. Only a streak and some RNA like band is seen far below in the gel..can uplease tell me why iam not able to get the DNA band.. I used the plasmid mini prep methof for the DNA isolation
Reply With Quote
  #2  
Old 06-09-2009, 03:06 PM
danfive's Avatar
Nobel Laureate
Points: 8,612, Level: 64 Points: 8,612, Level: 64 Points: 8,612, Level: 64
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2007
Location: Houston TX
Posts: 969
Thanks: 12
Thanked 184 Times in 154 Posts
Default Re: Why iam not able to get a band from my Plasmid DNA samples?

Did you use a mini prep kit? I don't know if you refer to cell pellet or DNA pellet.

My best advice for now is to check the DNA concentration OD 260/280.
You don't want to overload the agarose gel.
Reply With Quote
  #3  
Old 06-11-2009, 11:14 AM
Pipette Filler
Points: 22, Level: 1 Points: 22, Level: 1 Points: 22, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Why iam not able to get a band from my Plasmid DNA samples?

Yes. Check the DNA concentration and also what kind of gel are you running? I found that 0,8 % agarose gel in TEA buffer run at about 60-70 V to be best for viewing plasmids
Reply With Quote
  #4  
Old 03-28-2013, 06:47 AM
Pipette Filler
Points: 52, Level: 1 Points: 52, Level: 1 Points: 52, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2013
Posts: 7
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Why iam not able to get a band from my Plasmid DNA samples?

Check nanodrop reading of your elute, then analyse the plasmid using 0.8% agarose gel.
Reply With Quote
Reply

Tags
band , dna , iam , plasmid , samples , _


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
problem with pWATERGATE plasmid ismawanto Molecular Cloning Forum 5 05-31-2010 10:30 AM
YEp as a multi copy plasmid DB_Roland Yeast Forum 0 11-27-2008 01:44 PM
Plasmid Preps from MRSA Anthony C. Hilton Protocols and Methods Forum 1 07-10-2004 12:04 AM
yeast plasmid preps johnspar@earthlink.net Yeast Forum 0 02-20-2004 02:13 AM
Yeast plasmid preps Timothy Caspar Protocols and Methods Forum 0 01-30-2004 06:26 PM


All times are GMT. The time now is 04:07 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 0.12537 seconds with 16 queries