cy3 and cy5-dCTP labelling

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
bsengez
Garter
Garter
Posts: 10
Joined: Wed Aug 29, 2007 1:55 pm

cy3 and cy5-dCTP labelling

Post by bsengez » Thu Jul 22, 2010 12:54 pm

I purified total RNA from two different cell lines stably expressing an empty vector (as a control) and vector having two inserts. Now, I want to perform microarray. We have Cy3-dCTP and CY5-dCTP. Some protocols want you to purify mRNA and reverse transcribe it to cDNA, the others use total RNA to make cDNA. Which one is more suitable for these dyes? Is anyone using these dyes for cDNA or RNA labelling? I need a good protocol for it.
Thanks in advance..
Burcu

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Thu Jul 22, 2010 6:34 pm

IMHO that doesn't matter. Just in case of total RNA you must use higher amount of RNA, as the mRNA is only small fraction
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

bsengez
Garter
Garter
Posts: 10
Joined: Wed Aug 29, 2007 1:55 pm

Post by bsengez » Thu Jul 22, 2010 6:42 pm

How much RNA do I need to start with? Do you have any protocols for cDNA labelling?

Post Reply

Who is online

Users browsing this forum: No registered users and 3 guests