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Summary: Nucleotide exchange and excision technology
(NExT) DNA shuffling: a robust method for
DNA fragmentation and directed evolution
Kristian M. Mu¨ller*, Sabine C. Stebel, Susanne Knall, Gregor Zipf1
,
Hubert S. Bernauer1
and Katja M. Arndt
Institut fu¨r Biologie III, Universita¨t Freiburg, Scha¨nzlestraße 1, 79104 Freiburg, Germany and
1
ATG:Biosynthetics, Freiburg, Germany
Received March 30, 2005; Revised and Accepted July 8, 2005
ABSTRACT
DNA shuffling is widely used for optimizing complex
properties contained within DNA and proteins.
Demonstrated here is the amplification of a gene
library by PCR using uridine triphosphate (dUTP) as
a fragmentation defining exchange nucleotide with
thymidine, together with the three other nucleotides.
The incorporated uracil bases were excised using
uracil-DNA-glycosylase and the DNA backbone sub-
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