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Methods Mol Biol. 2009;498:75-90. doi: 10.1007/978-1-59745-196-3_5.

The precise engineering of expression vectors using high-throughput In-Fusion PCR cloning.

Author information

1
Oxford Protein Production Facility, Welcome Trust Centre for Human Genetics, Oxford, UK.

Abstract

In this chapter, protocols for the construction of expression vectors using In-Fusion PCR cloning are presented. The method enables vector and insert DNA sequences to be seamlessly joined in a ligation-independent reaction. This property of the In-Fusion process has been exploited in the design of a suite of multi-host compatible vectors for the expression of proteins with precisely engineered His-tags. Vector preparation, PCR amplification of the sequence to be cloned and the procedure for inserting the PCR product into the vector by In-Fusion are described.

PMID:
18988019
DOI:
10.1007/978-1-59745-196-3_5
[Indexed for MEDLINE]

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