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[Establishment and preliminary application of a quantitative real-time PCR method for detecting the transcriptions of Th1/Th2 cytokines in RAW264.7 cells].

Authors :
Shan X
Han X
Zhang M
Song J
Liu H
Tian M
Pan L
Ding C
Zhou J
Yu S
Source :
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2013 Apr; Vol. 29 (4), pp. 430-3.
Publication Year :
2013

Abstract

Objective: To establish a quantitative real-time PCR method to detect the transcriptions of Th1/Th2 cytokines in RAW264.7 cells.<br />Methods: The specific primers were designed according to the sequences of TNF-α, IFN-γ, IL-2, IL-4, IL-6, IL-10 and β-actin in GenBank database. Total RNA was extracted from RAW264.7 cells, and then was reverse-transcriped into cDNA, which was established as a positive standard template of the quantitative real-time PCR method. The established method was used to detect the cytokine transcriptions in RAW264.7 cells infected with Brucella abortus S2308.<br />Results: Cytokine genes above had a good linear relationship (R(2);≥ 0.982) with the detection limit of 10(2); copies/μL standard samples. The established quantitative real-time PCR method showed high specificity, sensitivity, and single melting peak for every cytokine.<br />Conclusion: The quantitative real-time PCR method for detecting the transcription levels of cytokine Th1 and Th2 of macrophage RAW264.7 cells was established successfully.

Details

Language :
Chinese
ISSN :
1007-8738
Volume :
29
Issue :
4
Database :
MEDLINE
Journal :
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Publication Type :
Academic Journal
Accession number :
23643175