high percentage agarose

Dr Omar Hamarsheh via methods%40net.bio.net (by ohamarsheh from gmail.com)
Mon Aug 11 14:38:45 EST 2008


Dear Rashmi,

I suggest on you to use MetaPhore agarose, 4.5 % will be very good for small
fragments. Be aware that you need to cook it carefully. Fist use cold buffer
(stored on 4C), add the MetaPhore garose powder little by little while
mixing using magnetic stirrer, let the mix for 15 minutes then cook it in
the microwave starting at low heat then increase the temperature gradually
(it needs 20- 30 min ).

For running the gel, 150 watts for 4-5 hours, do not put stain (Ethedium
bromide) with the gel, just incubate the gel after running for 15-20 min in
a bath of Eth. Brd diluted with distilled water 

All the best,
Omar   

-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of rashmi
Sent: Monday, August 11, 2008 6:31 PM
To: methods from magpie.bio.indiana.edu
Cc: rashmi21481 from gmail.com
Subject: high percentage agarose

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
AIIMS
New Delhi
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