Quantification by real-time PCR of developmental and adult myosin mRNA in rat muscles.

Abstract:

:A real-time RT-PCR assay using newly designed primers was developed to analyze developmental and adult MHC mRNA expression both in skeletal muscles and single fibers. Only 4 ng of total RNA was necessary for the analysis of the relative mRNA expression of MHC genes. Different validation steps were realized concerning both specificity and sensitivity of each primer set, and linearity and efficiency of each real-time PCR amplification. Then, quantification of MHC mRNA in neonatal and adult muscles as well as in single fibers was done by the deltaC(T) method, with CycA gene as the reference gene. Due to a higher sensitivity than that of a competitive PCR method, we demonstrated that this assay is suitable to study very low level of MHC mRNA expression as developmental MHC in adult muscle and to quantify mRNA from very small samples.

authors

Sanchez H,Chapot R,Banzet S,Koulmann N,Birot O,Bigard AX,Peinnequin A

doi

10.1016/j.bbrc.2005.11.172

subject

Has Abstract

pub_date

2006-02-03 00:00:00

pages

165-74

issue

1

eissn

0006-291X

issn

1090-2104

pii

S0006-291X(05)02700-2

journal_volume

340

pub_type

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