How necessary to prevent self-ligation using CIP?

[Helge Weissig]

In article <2sfak3$msl at kantti.Helsinki.FI>, zheng at cc.Helsinki.FI (Huanquan
Zheng) wrote:

> 
> Yes, to prevent recircularization of lineared vector, it is abselutely
> needed.
> 
> But when you guys do the cloning, in the following cases, is it
> absolutely necessary to use CIP to prevent self-ligation of vertor?
> 
> 1. vector digested with BamH I & Kpn I with which insert will be digested too.

no need for CIP here as generally for all non-compatible double-digests.

> 2. blunt end vector (which blunt end insert will be ligated into)

just as with sticky ends, you want to CIP-treat ("zip") the vector to
prevent circulization. I usually include CIP with the restriction enzyme.

> 
> 
> --
> 	Huanquan Zheng 	~{V#;7H*#,KD4(HK#,;lHUWS5D#,~} 
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hope this helps,
cheers
Helge

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Helge WEISSIG                              e-mail: helgew at ljcrf.edu
La Jolla Cancer Research Foundation        phone: (619) 455 6480 x253
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