Chimera construction using multiple-template-based sequential PCRs.

Abstract:

:Chimera construction between different proteins is a useful method for investigating protein structure and function relationships. However, this technique, by its traditional application, has been daunting because of its complex procedure and low success rate. Here we describe a protocol for constructing chimeras between proteins that does not require the existence of restriction sites, or the purification of intermediate PCR products, which are essential in the traditional protocols. By introducing the "multiple-template" concept, this protocol only requires the use of two or three simple PCRs followed by general subcloning steps. Most importantly, the success rate is nearly 100%.

journal_name

J Neurosci Methods

authors

Shan Q,Lynch JW

doi

10.1016/j.jneumeth.2010.08.033

subject

Has Abstract

pub_date

2010-10-30 00:00:00

pages

86-9

issue

1

eissn

0165-0270

issn

1872-678X

pii

S0165-0270(10)00493-0

journal_volume

193

pub_type

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