title: [Establishment and preliminary application of a quantitative real-time PCR method for detecting the transcriptions of Th1/Th2 cytokines in RAW264.7 cells].
abstract: OBJECTIVE
  To establish a quantitative real-time PCR method to detect the transcriptions of Th1/Th2 cytokines in RAW264.7 cells.
  
  
  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.
  
  
  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.
  
  
  CONCLUSIONS
  The quantitative real-time PCR method for detecting the transcription levels of cytokine Th1 and Th2 of macrophage RAW264.7 cells was established successfully.
authors: School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China. sxq06113071@163.com
authors: Xueqin
authors: X
authors: Shan
authors: Xiangan
authors: X
authors: Han
authors: Min
authors: M
authors: Zhang
authors: Jun
authors: J
authors: Song
authors: Haiwen
authors: H
authors: Liu
authors: Mingxing
authors: M
authors: Tian
authors: Ling
authors: L
authors: Pan
authors: Chan
authors: C
authors: Ding
authors: Jinping
authors: J
authors: Zhou
authors: Shengqing
authors: S
authors: Yu
mesh_terms: D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D051379:Mice; D012313:RNA / Q000032:analysis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018417:Th1 Cells / Q000276:immunology* / Q000378:metabolism; D018418:Th2 Cells / Q000276:immunology* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D016207:Cytokines; D018076:DNA, Complementary; D012313:RNA