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[Constructing a Competitor of BCR-ABL cDNA by PCR Site-Directed Mutagenesis]
Hong Tian1, Jing Sun, Xiaomian Zhou
1Department of Hematology, Department of Docimasiology, Nanfang Hospital, Guangzhou 510515, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi
|February 13, 2003
Abstract:
A competitive mimic of the cDNA of the BCR-ABL fusion gene was constructed, and its feasibility was testified by capillary electropheresis (CE). The 4 bp-shorter mimic was obtained by PCR amplification using a newly synthesized downstream primer analogous to the former one. Mimics of both types of BCR-ABL cDNA were achieved and the validity was verified with restriction endonuclease. And the products of the coamplification PCR could be easily separated by capillary electrophorisis. The mimic can be used to quantitative detection of BCR-ABL gene through competitive RT-PCR in chronic myeloid leukemia.