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

Electrophoretic Mobility Shift Assay Forum EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


major Issues with EMSA!! Please HELP!!!!!!

major Issues with EMSA!! Please HELP!!!!!! - Electrophoretic Mobility Shift Assay Forum

major Issues with EMSA!! Please HELP!!!!!! - EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 11-07-2008, 10:02 PM
Pipette Filler
Points: 158, Level: 3 Points: 158, Level: 3 Points: 158, Level: 3
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Unhappy major Issues with EMSA!! Please HELP!!!!!!



I am running EMSA assays with my DNA binding protein and a 94 base pair oligo. I am doing a gradient where I keep the DNA conc same but increase the protein conc. I am using low salt binding buffer and MGcl2 in the mix. I am running it in a 4% PAGE gel. However all I am getting in addition to really bright bands of unbound DNA, are smears in all the lanes. and these smears keep decreasing in intensity as the protein conc increases. I dont know is thats binding or some artifact..but its consistent from gel to gel. I really need help solving this issue .....so please please help me out!
Reply With Quote
Reply

Tags
dna binding , emsa , gel electrophoresis , issues , major , protein:dna binding


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
Rna Emsa tara29466 RNA Techniques Forum 4 06-29-2011 10:35 AM
EMSA with Proteins which bind non-specifically to DNA Adelaide_Science Electrophoretic Mobility Shift Assay Forum 2 12-10-2008 02:09 PM
Problem with EMSA - Undefined bands amazonic9 Protocols and Methods Forum 11 01-10-2008 01:11 PM
EMSA problem Balat Protocols and Methods Forum 1 11-20-2006 08:39 AM
EMSA help...please URGENT sumo1 Protocols and Methods Forum 5 09-14-2006 09:50 AM


All times are GMT. The time now is 06:14 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 0.11083 seconds with 16 queries