high percentage agarose

Dunowska, Magda via methods%40net.bio.net (by M.Dunowska from massey.ac.nz)
Mon Aug 11 19:57:17 EST 2008

A long time ago I was looking for small differences between fragments and used pre-cast polyacrylamide gels (from Bio-Rad I think), which I stained with a gel-star nucleic acid stain (currently sold by Lonza). Worked really well - Magda

Magda Dunowska, LW (vet), PhD
Senior Lecturer in Veterinary Infectious Diseases (Virology)
Institute of Veterinary, Animal and Biomedical Sciences
Te Kura Mâtauranga Kararehe
Massey University
Palmerston North
New Zealand

Phone : (06) 356-9099 ext 7571
Website : http://ivabs.massey.ac.nz
-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu [mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Dr. Hiranya S. Roychowdhury
Sent: Tuesday, 12 August 2008 9:22 a.m.
To: rashmi
Cc: methods from magpie.bio.indiana.edu; rashmi21481 from gmail.com
Subject: Re: high percentage agarose

The opening line always fascinates me!  The email was directed to the
methods listserve, and to be sure, there would be someone in this group wo
had posted on some unspecified "google groups"!

Anyway, I suggest a mini acrylamide sequencing set-up. There are many
protocols for that in places as you have mentioned.

> Hi there,
> I am a PhD student working on Pneumocystis biodiversity. I cam across
> your message posted on google groups, regarding high percentage
> agarose gels and TBE concentration. One of the objective of my thesis
> is to do typing by counting number of 10 base pair repeats in my PCR
> products. The number of repeats can vary from 2 to 6. The sizes are
> 125, 135, 145, 155 and 165. I tried to run a 4% agarose gel on a tray
> about 25 cm. But my gel was not properly formed. i will be highly
> delighted if you could suggest me something, like the % of agarose I
> should use, run time, gel lengh, voltage etc.
> Thanks in advance
> Rashmi Gupta
> PhD Student
> New Delhi
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Hiranya S. Roychowdhury, Ph.D.
Asst. Professor,
Health & Public Services
Dona Ana Community College
New Mexico State University
Las Cruces, NM 88003

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