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Genotype identification of Math1/LacZ knockout mice based on real-time PCR with SYBR Green I dye.

Abstract：

:Knockout mice are widely used in all fields of biomedical research. Determining the genotype of every newborn mouse is a tedious task, usually performed by Southern blot hybridization or Polymerase Chain Reaction (PCR). We describe here a quick and simple genotype identification assay based on real-time PCR and SYBR Green I dye, without using fluorescent primers. The discrimination between the wild type and targeted alleles is based on a PCR design that leads to a different melting temperature for each product. The identification of the genotype is obvious immediately after amplification, and no post-PCR manipulations are needed, reducing cost and time. Therefore, while the real-time PCR amplification increases the sensitivity, the fact that the reactions tubes are never opened after amplification, reduces the risk of contamination and eliminates errors, which are common during the repeated handling of dozens of samples from the same mouse line. The protocol we provide was tested on Math1 knockout mice, but is general, and may be utilized for any knockout line and real-time thermocycler, without any further modification, accessories or special reagents.

journal_name

J Neurosci Methods

journal_title

Journal of neuroscience methods

authors

Krizhanovsky V,Golenser E,Ben-Arie N

doi

10.1016/j.jneumeth.2004.01.007

subject

Has Abstract

pub_date

2004-07-30 00:00:00

pages

187-92

issue

2

eissn

0165-0270

issn

1872-678X

pii

S0165027004000287

journal_volume

136

pub_type

杂志文章

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