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30 October 2012, Volume 29 Issue 05

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A Simple and Efficient Relative Quantitative Method of FQ-PCR

CHEN Jun, WANG Yin, LI Hui-Ling, DONG Jian-Yi, WANG Fu-Jin, WANG Ai-Guo, WANG Jing-Yu

2012, 29(05):  1-5. 

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Abstract: Objective To establish a simple and efficient relative quantitative method of FQ-PCR． Method Specific primers for Fabp5，Ppar-α and β-Actin with strict and consensus parameters were designed，and the corresponding plasmid standards were prepared to establish the standard curves after 10-fold dilution． Then the three standard curves were used to quantify the differential expression of Fabp5，Ppar-α and β-Actin in the liver tissue of normal mice，the liver tumor and peri-tumor tissues of H-ras12V transgenic mice by single-standard curve， 2 － △△Ct and double-standard curve methods，respectively． β-Actin was used as an internal control． Result There was no significant difference in the relative quantitative value determined by single-standard curve method and by double-standard curve method． However，compared to double-standard curve method，relative quantitative value determined by 2 － △△Ct method fluctuated largely． Conclusion Single-standard curve method could be applied for relatively quantitative analysis of differentially expressed gene based on the specific primers designed with strict and consensus parameters．