How to remove a possible hairpin from a vector

j dsfjo santbrin at CHEM.LEIDENUNIV.NL
Wed Dec 10 08:14:22 EST 1997


After ligating a linker with an internal Bam HI and Not I site in a
vector, I'am not able to linearize the vector with not I.
The presence of the linker is confirmed by digestion with other enzymes.
Since the Not I site consists of only G's and C's and the vector also
has a few G's and C's at the Not I recognision site I suspect somehow
the formation of a hairpin loop which destroys the Not I recognision
site.
Is this a possibility and if so, what can I do about it (DMSO, betaine
formamide) ?

any help is appreciated.

peter van santbrink
division of Biopharmaceutics
Leiden University
the Netherlands



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