Help with bands resolution

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

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
Posts: 2
Joined: Wed Mar 23, 2011 4:55 pm

Help with bands resolution

Post by Dagon123 » Wed Apr 20, 2011 1:57 am


I'm trying realize a subcloning. First, i digested 2ug of the vector with EcoRI to release the insert, the insert size is about 5,5kb and the backbone vector i'ts about 6,8Kb. When i runned the digestion into an agarose gel i saw only one band, and i need to purify only the 5,5kb band.
I'm using 1% of agarose concentration and running the gel to 100V for 2 hours. I also merge two wells of the gel to load the 2ug of digested DNA.

My question is, what is the correct agarose concentration and voltage that i have to use to resolve both bands? maybe i need to use lower agarose concentration and lower voltage too.. but,, what is the correct procedure?.

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

Post by JackBean » Wed Apr 20, 2011 5:18 am

0.7% or 0.8% gel should be fine

Cis or trans? That's what matters.

Post Reply

Who is online

Users browsing this forum: Google Adsense [Bot] and 7 guests