Protocol Online logo
Top : Forum Archives: : Molecular Biology

Tool for complementary restriction enzymes? - REs producing overhangs to religate (Jun/23/2008 )

I am looking for a tool or a webpage, where I can find complementary REs.
For example I want cut a gene with 2 RE and religate the vector so that the gene is removed but the vector is circular again and the reading frame for proteinexpression has not changed.

my 3´ RE is KpnI. Now I am looking for a RE that generates a complementary overhang that can be ligated with a KpnI overhang.

Genestructure:

RE1 - gene1 - KpnI - gene2 - RE3

gene2 was cloned via KpnI and RE3. Now I want to design the vector in that way, that optionally I can get rid of gene1 by digesting with RE1 and KpnI and religate the vector so that the frame stays intact.

RE1 must not be KpnI as KpnI is also used for cloning gene2 into the vector.

understandable? ;-)

Is there a tool or a webpage, giving such information for any RE?

THX

-Dukes-

Try this site

http://www.neb.com/nebecomm/tech_reference...e_overhangs.asp

-smu2-

if you are running a mac - EnzymeX is a good freeware.

-bob1-

Thank you for your help.

I somehow have missed this information at NEB.... probably I am getting old...;-)

As KpnI doesn´t seem to have such a "partner", I will use Acc65I (isoschizomere of KpnI) and BsiWI instead.

-Dukes-

Hallo type REBASE in the google and in this databese search feature choose recognition sequence and type KpnI sequence
or design primers with 5 prime overhang with whatever RS
good luck

-baxapoptoaia-

CLC free workbench is also good and it has a large catalog of REs. VectorNTI is a very nice software, you can do basically anything you want with it, even contig assembly. it's freeware but you must register

-toejam-