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

RNA Techniques Forum Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols.


RNA integrity - strange bands on the gels

RNA integrity - strange bands on the gels - RNA Techniques Forum

RNA integrity - strange bands on the gels - Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 07-30-2007, 02:02 PM
Luisinlab's Avatar
Pipette Filler
Points: 1,117, Level: 19 Points: 1,117, Level: 19 Points: 1,117, Level: 19
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2007
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Question RNA integrity - strange bands on the gels



I run some samples from plant tissues side-by-side on a non-denaturing 2% agarose gel (in TBE) and I have some samples with higher-weight ribossomal bands than others. The intensity ratio is OK in every sample and there are no smears. What is happening? (The samples are from similar tissues from the same species!!)

Help me please. I need to know if some of this RNA is not suitable for RT-PCR!!!

see attached picture (line 1 -ladder; lane 2 (suspected sample)
What is the difference between line 3 and lines 4-8???
Attached Images
File Type: jpg RNA.jpg (32.2 KB, 82 views)

Last edited by Luisinlab; 07-30-2007 at 02:13 PM. Reason: to add a picture file
Reply With Quote
  #2  
Old 07-30-2007, 04:00 PM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: RNA integrity - strange bands on the gels



RNA GEL

Hello there and welcome to the forums!
My guesses in no particular order:
1) sample contaminations
2) salt differences
3) loading amount differences

if you are sure you didn't contaminate your samples then did you extract them all the same way at the same time?

please tell us what you found
cheers!

Last edited by admin; 07-30-2007 at 04:03 PM.
Reply With Quote
  #3  
Old 07-31-2007, 04:12 PM
Luisinlab's Avatar
Pipette Filler
Points: 1,117, Level: 19 Points: 1,117, Level: 19 Points: 1,117, Level: 19
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2007
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: RNA integrity - strange bands on the gels

They were not extracted at the same time but the solutions were the same...
Salt differences? What do you mean by that? (different concentrations; for sure! but that explains the lower weight of the bands?)
Reply With Quote
  #4  
Old 08-01-2007, 11:34 AM
Pipette Filler
Points: 4,137, Level: 43 Points: 4,137, Level: 43 Points: 4,137, Level: 43
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2007
Posts: 5
Thanks: 0
Thanked 1 Time in 1 Post
Default Re: RNA integrity - strange bands on the gels

Hi

How did you store the samples between isolations, how were you preparing the tissue for isolation, was it unfrozen for a longer period of time ...? RNA is not as stable as some of us would like it to be when we are working with it, so it could be that you didn't handle the samples correctly and it was partialy degraded. If you are 100% sure that you can exclude contamination (you might have had something in your buffers from 1st extraction on) this might explain what is going on.

neja

Last edited by neja; 08-01-2007 at 11:41 AM.
Reply With Quote
  #5  
Old 08-01-2007, 01:14 PM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: RNA integrity - strange bands on the gels

Quote:
Originally Posted by Luisinlab View Post
They were not extracted at the same time but the solutions were the same...
Salt differences? What do you mean by that? (different concentrations; for sure! but that explains the lower weight of the bands?)
Yes salt differences can make large differences in the migration of RNA or DNA in agarose gels. This may have been your problem.

Did you purify samples or desalt them prior to running them?

Reply With Quote
  #6  
Old 10-07-2009, 10:03 PM
Pipette Filler
Points: 62, Level: 1 Points: 62, Level: 1 Points: 62, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Oct 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: RNA integrity - strange bands on the gels

I've ran my samples from fish and I've got three bands, two more or less normal and a third one strange on the top. Do you know what can it be?
Reply With Quote
Reply

Tags
bands , gels , integrity , rna , strange


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
Long Term Storage of SDS-PAGE Gels dnamolecule SDS-PAGE Gel Electrophoresis Forum 1 08-28-2008 06:34 AM
Immidiate dissappearing of bands on DNA sequencing gels after silver staining himanshu SDS-PAGE Gel Electrophoresis Forum 2 07-24-2008 06:17 PM
Increase Exposure, Bands Disappear?? Mediatorx Western Blot Forum 3 07-01-2008 09:18 PM
Help Migrating bands camherst Western Blot Forum 1 01-26-2008 02:48 PM
qPCR NEWSLETTER September 2006 Editor www.Gene-Quantification.info Protocols and Methods Forum 0 09-28-2006 12:43 PM


All times are GMT. The time now is 02:39 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.14457 seconds with 17 queries