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

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

Re:purification of radioactive probe

Re:purification of radioactive probe - Protocols and Methods Forum

Re:purification of radioactive probe - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.

LinkBack Thread Tools Display Modes
Old 08-27-2007, 11:14 PM
Analia Alet
Posts: n/a
Default Re:purification of radioactive probe

Hi Simone,

I used to make DNA probes for northern blot by using hexamers and the Klenow
fragment in the reaction mix. Anyway, after the reaction was finished, I
purified it with a Sephadex G-50 column. The reason I did so was because
according to Manniatis, Sephadex G-50 retains the unused labelled-nucleotides
and little oligonucleotides that may interfere in the blotting. After using
the column, the eluate was cheked for radiactivity, boiled 10 min, and
transferred to the hybridisation solution.

Once againg, please forgive me my english
Best luck

Lic. Analía I. Alet
Unidad de Biotecnología 1
Camino Circunvalación Laguna km 6
Pcia de Buenos Aires
Tel: 0054 2241 430323 int102


does anyone know if it is really necessary to purify a radioactive DNA probe
after labelling it by random priming? I want to use it on a northern blot.
So far, I worked with DIG-labelled probes, that do not need to be purified
after labelling. They are used directly in the hybridisation solution. Is it
different with radioactive probes? Do you think that precipitation by salt
and alcohol is sufficient for purification?

Thank you very much,

Reply With Quote

probe , radioactive , repurification

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
Real Time-PCR probe design Ganto Real-Time PCR and Quantitative PCR Forum 1 09-17-2009 11:24 AM
minimum radioactive probe size Ed Siefker Protocols and Methods Forum 1 09-26-2007 08:50 PM
purification of radioactive probe Simone Marker Protocols and Methods Forum 2 08-28-2007 12:13 PM
purification of radioactive probe Jayakumar, R Protocols and Methods Forum 0 08-27-2007 04:48 PM
problems with probe binding to 28S and 18S RNA only on Northerns Wendy Ingram Protocols and Methods Forum 0 09-11-2006 02:30 AM

All times are GMT. The time now is 07:22 AM.

Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2015, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11994 seconds with 16 queries