PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of narLink to Publisher's site
 
Nucleic Acids Res. Oct 15, 1997; 25(20): 4165–4166.
PMCID: PMC147003

Ligation independent cloning irrespective of restriction site compatibility.

Abstract

Here we report the use of exonuclease to expose complementary DNA between an insert and vector such that annealing becomes independent of restriction site compatibility. We demonstrate that unusual and, in some cases, previously impossible cloning strategies can be readily and efficiently achieved as long as the flanking sequences of the linear vectors are highly related. Furthermore, we show that the bacterial repair system resolves the residual mismatches, overhangs or gaps in a predictable fashion to generate excisable inserts. This approach facilitates cloning regardless of restriction site compatibility and overcomes an important limitation in current cloning techniques.

Full Text

The Full Text of this article is available as a PDF (56K).

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press