Question on Plasmid Cloning

no one no at one
Sun Aug 15 13:54:16 EST 1999


These are called concatamers and they are a problem.  They will ligate into
the vector just fine and the colonies will grow just fine.  The only way you
can spot them is to do a plasmid prep on each colony and run that on a gel.
If concatamers are there, they will run slower on the gel (i.e. the insert
will have a higher molecular weight).

Gary Krause
Wayne State University



Student <mucineer at iname.com> wrote in message
news:7oouq1$iha$1 at oyez.ccc.nottingham.ac.uk...
> Dear All
>
> As a new comer to molecular biology, I have the following trivial question
> and hope someone can help:
>
> When ligating a DNA insert into a plasmid (both the insert and plasmid
> contain different restriction sites on either ends, e.g. a
BamHI----HindIII
> fragment into a vector digested with these two restriction enzymes), is
> there anything in the process that prevent different molecule of the
insert
> from "self-ligating" to form species such as
> HindIII-------BamHI--------HindIII or BamHI--------HindIII------BamHI?
>
> This should be a common problem to all ligations but it doesn't seem to be
a
> big problem as I usually get hundreds of colonies on the plate.
> Does anyone have any insights into this?
>
> Any comments will be appreciated.
>
>
>
>
>
>





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