back, thought I'd give them a look and send off the ones with inserts.
Problem is, a majority of the clones have inserts of the same size, or
two sizes overall. My hope was, I could add to my data, possibly
finding a repetitive clone. In Gel 1, there are two clones that I think
have an insert (described below). In Gel 2, only 4 lanes have a
different insert size, or are blank. My guesses are: TOPO is ligating
to itself or some other phenomenon.
Pic 1:
Lane 1 is a kb ladder
Lane 2 in TOPO
Lane 3 is TOPO control cut with ecorI 750 bp insert
Lane 4-9 are TOPO vectors that are suppose to have inserts, selected
from white colonies. The inserts were made by randomly shearing genomic
DNA. Then ligated into TOPO. Lanes 4 and 7 have an insert. What about
the other lanes? They have an insert, but all are the same size.
Impossible. What is happening here.
Same with Pic 2:
Look at all the same sized inserts (of two various sizes). I count maybe
two real inserts, and some blank lanes. What is going on here?
Edited by relapse71, 20 August 2009 - 07:17 PM.