Biology-Online • View topic - Gel Electrophoresis Help!

Join for Free!
122503 members

Gel Electrophoresis Help!

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

Moderator: BioTeam

Gel Electrophoresis Help!

Postby cks123 » Tue Aug 27, 2013 4:25 pm

So I did a double digest for subcloning and I didn't get two distinct bands when I ran the gel. Can someone interpret the gel pattern for me?
pI-SceI site digest.jpg
(14.2 KiB) Not downloaded yet
Posts: 1
Joined: Tue Aug 27, 2013 4:19 pm

Postby JackBean » Wed Aug 28, 2013 2:11 pm

not really, because there is hardly something visible. What sizes is it? What sizes did you expect?

Cis or trans? That's what matters.
User avatar
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Postby bravebeaker » Thu Aug 29, 2013 7:14 am

size of your vector/insert.
gel is prestained with ethidium bromide or not.
what are the restriction enzymes you are using?
did you check if your vector contains multiple sites of the enzymes you are using?

The bands are way up, higher than 10k bp..
AHH, Gravity! Such a cold cruel mistress.
User avatar
Posts: 44
Joined: Sun May 08, 2011 3:26 am
Location: Kobe, Japan

Postby Cat » Sun Sep 01, 2013 2:23 pm

It looks like a bad gel. I suggest to re-run with fresh buffer/gel.
King Cobra
King Cobra
Posts: 635
Joined: Thu Feb 14, 2008 7:40 pm

Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 1 guest