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

RNA Techniques Forum Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols.

Tail of the RNA band

Tail of the RNA band - RNA Techniques Forum

Tail of the RNA band - Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols.

LinkBack Thread Tools Display Modes
Old 04-03-2008, 10:11 PM
Pipette Filler
Points: 626, Level: 12 Points: 626, Level: 12 Points: 626, Level: 12
Activity: 0% Activity: 0% Activity: 0%
Join Date: Mar 2008
Posts: 2
Thanks: 0
Thanked 0 Times in 0 Posts
Unhappy Tail of the RNA band

Hi guys,

I am a chemistry graduate student. Currently I am transcripting a 50 bp long RNA from 70 bp long DNA. I am using the Ampliscribe T7 flash transcription Kits.
When I check the purity of purified RNA ( purified by cutting the 10% denaturing PAGE gel in TBE buffer) in 20% denaturing PAGE gel. I always got a significant tail of the RNA band ( which is in the lower mobility direction).

I am pretty sure that I have cutted the purification gel correctly. There should be no other impurities (such as template DNA, which has been digested by RNase free DNase I).

Could you please tell me what might be the reason and how can I get clean RNA product?

Reply With Quote
Old 04-09-2008, 10:50 PM
admin's Avatar
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: Tail of the RNA band

Hello there DNA Artist,
there could be several reasons.

THe first is that you somehow have a longer DNA. Either by not cutting fully or by PCR mistakes (it is longer than 70bp).

The other reason is that you have degradation, meaning that you get a smear instead of a clear band.

To get a clean RNA product, I would make sure you cut the DNA 100% to completion to have runoff transcripts and also use a sephadex column to get rid of nucleotides and small unfinished (but partly transcribed) RNAs.

I used RNAse free-Sephadex G-25 for less than 200bp RNAs (Roche carries them).

please let me know how you are doing.

Reply With Quote

band , rna , tail

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
iritating band mollizi Western Blot Forum 0 02-03-2010 02:31 PM
[help] the tail of RNA band DNAartist RNA Techniques Forum 0 03-28-2008 07:56 PM
white sample band but black positive control band wilairat Western Blot Forum 6 01-27-2008 01:13 AM
band separation prob. on agarose gels Kyle Legate Protocols and Methods Forum 5 02-27-2004 10:24 PM
combining Conduction Band theory with Electronegativity as one phenomenon Archimedes Plutonium Chemistry Forum 2 10-12-2003 06:25 PM

All times are GMT. The time now is 05:29 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 0.11761 seconds with 16 queries