RRC ID 1890
Author Nour-Eldin HH, Hansen BG, Nørholm MH, Jensen JK, Halkier BA.
Title Advancing uracil-excision based cloning towards an ideal technique for cloning PCR fragments.
Journal Nucleic Acids Res
Abstract The largely unused uracil-excision molecular cloning technique has excellent features in most aspects compared to other modern cloning techniques. Its application has, however, been hampered by incompatibility with proof-reading DNA polymerases. We have advanced the technique by identifying PfuCx as a compatible proof-reading DNA polymerase and by developing an improved vector design strategy. The original features of the technique, namely simplicity, speed, high efficiency and low cost are thus combined with high fidelity as well as a transparent, simple and flexible vector design. A comprehensive set of vectors has been constructed covering a wide range of different applications and their functionality has been confirmed.
Volume 34(18)
Pages e122
Published 2006-1-1
DOI 10.1093/nar/gkl635
PII gkl635
PMID 17000637
PMC PMC1635280
MeSH Cloning, Molecular / methods* DNA-Directed DNA Polymerase / metabolism Deoxyribonucleases, Type II Site-Specific Genetic Vectors Polymerase Chain Reaction* Uracil / metabolism*
IF 11.502
Times Cited 297
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Arabidopsis / Cultured plant cells, genes pda05545 pda03530 pda01560 pda04389 pda04574