Fw: Fw: What is the effect of a 5' overhang on gel mobility?

Deanne Bell dbell at qnis.net
Thu Jan 11 11:29:16 EST 2001


To Dr. Hiranya S. Roychowdhury"
I must comment on how polite you are!!  Rather than being so rude, would
you kindly tell me where my logic is incorrect?
I made it quite clear that I have not done Restriction fragment gels
before.  And the original question did not clearly state their puposes.
D. Bell

----------
> From: "Dr. Hiranya S. Roychowdhury" <hroychow at nmsu.edu>
> To: methods at hgmp.mrc.ac.uk
> Subject: Re: Fw: What is the effect of a 5' overhang on gel mobility?
> Date: Wednesday, January 10, 2001 6:31 PM
> 
> Do you really know what you are talking about?
> >
> >
> >I haven't really done any of this work but i would think that 4 bp would
> >make a difference.
> >When you run restriction fragments out on a gel, I think the fragments
are
> >denatured and run in such a manner that you have resolution to 1 - 2 bp.
> >Kind of like a sequencing gel.  In my humble opinion (IMHO) if you want
> >these two fragments to migrate down a gel at the same speed, you should
> >make the PCR product as long as the restriction fragment including the
> >overhang.  Then denature all the fragments and run the gel.
> >
> >Deanne Bell
> >Molecular Markers Lab Technician
> >USDA Agricultural Research Service
> >2021 S. Peach Ave.
> >Fresno, CA 93727
> >e-mail: dbell at qnis.net
> >phone: 559 453-3170
> >fax: 559 453-3088
> >
> >
> >---
> >
> >
> >
> Dr. Hiranya Sankar Roychowdhury
> College Asst. Prof.
> Molecular Biology,
> Dept. of Chemistry & Biochemistry	
> Box 30001 - 3MLS
> New Mexico State University
> Las Cruces, NM 88003
> 
> Lab: (505) 646 4722
> Office: (505) 646 8256
> hroychow at nmsu.edu
> 
> 
> ---
> 
> 


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