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  • Title: [Real-time quantitative RT-PCR for the detection of bcl-2 mRNA expression].
    Author: Zhang Z, Lu L, Lan X, Hu J, Lin M, Jing Q.
    Journal: Zhonghua Yi Xue Yi Chuan Xue Za Zhi; 2002 Oct; 19(5):412-5. PubMed ID: 12362319.
    Abstract:
    OBJECTIVE: To establish a real-time quantitative RT-PCR method for the detection of expression of bcl-2 mRNA. METHODS: The vector containing bcl-2 gene as standard template was constructed with T-A cloning technique. The fluorogenic probe (i.e.,Taq man probe) was used to establish a real-time RT-PCR. bcl-2 mRNA expression level in Burkitt's lymphoma pre-and post-treated with bcl-2 antisense phosphothioate oligodeoxynucleotides (AS-PS-ODN) was determined with real-time quantitative RT-PCR, also with semi-quantitative RT-PCR. RESULTS: The expression of bcl-2 mRNA in Burkitt's lymphoma treated with bcl-2 AS- PS-ODN decreased significantly and no changes of bcl-2-mRNA expression in group treated with nonsense ODN were noticed. The semi-quantitative RT-PCR results also demonstrated that bcl-2 level varied as detected with real-time fluorogenic quantitative RT-PCR, but less sensitive and accurate. CONCLUSION: Detection of bcl-2 mRNA expression with the fluorogenic real-time quantitative RT-PCR is more accurate and sensitive than semi-quantitive RT-PCR.
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