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2011•China Animal Husbandry & Veterinary MedicineRequires access

Establishment of Real-time PCR for Mouse β-actin mRNA

Cheng Tang

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Abstract

The aim of present study is to establish a real-time PCR for quantitative analysis of mouse β-actin mRNA expression.A set of primer was designed according to the mouse β-actin mRNA sequence in GenBank.And a SYBR Green Ⅰ-based reverse real-time PCR(RRT-PCR) was successfully developed.The results showed that RRT-PCR assay established had a high amplification efficiency(97.7%),wide linear range(9.3×102 to 9.3×107 copies/reaction),good reproducibility and specificity.The detection limit was 9.3 copies/reaction.The results provided a basis for use of β-actin as a reference gene in gene expression analysis in mouse.

About this research paper

What this paper is about

The aim of present study is to establish a real-time PCR for quantitative analysis of mouse β-actin mRNA expression.A set of primer was designed according to the mouse β-actin mRNA sequence in GenBank.And a SYBR Green Ⅰ-based reverse real-time PCR(RRT-PCR) was successfully developed.The results showed that RRT-PCR assay established had a high amplification efficiency(97.7%),wide linear range(9.3×102 to 9.3×107 copies/reaction),good reproducibility and specificity.The detection limit was 9.3 copies/reaction.The results provided a basis for use of β-actin as a reference gene in gene expression analysis in mouse.

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Available abstract

The aim of present study is to establish a real-time PCR for quantitative analysis of mouse β-actin mRNA expression.A set of primer was designed according to the mouse β-actin mRNA sequence in GenBank.And a SYBR Green Ⅰ-based reverse real-time PCR(RRT-PCR) was successfully developed.The results showed that RRT-PCR assay established had a high amplification efficiency(97.7%),wide linear range(9.3×102 to 9.3×107 copies/reaction),good reproducibility and specificity.The detection limit was 9.3 copies/reaction.The results provided a basis for use of β-actin as a reference gene in gene expression analysis in mouse.

Key concepts: Real-time polymerase chain reaction, Primer (cosmetics), GenBank, Molecular biology, Biology, Messenger RNA, SYBR Green I, Actin

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