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本实验通过合成与原有下游引物 (B)相似的一条新引物 (B′) ,经聚合酶链反应 (polymerasechainreaction ,PCR)扩增出比原BCR ABLcDNA序列减少了 4个碱基的参照物 ,达到定点诱变的目的。经酶切分析证明 ,两型BCR ABLmRNA均可通过此方法得到相应的cDNA参照物。参照物和待测模板的共扩增产物可通过毛细管电泳达到基线分离 ,证明了其可行性。该参照物可用于慢性粒细胞白血病BCR ABL融合基因的竞争性PCR ,对此基因进行定量检测
In this experiment, a new primer (B′) similar to the original downstream primer (B) was synthesized, and a polymerase chain reaction (PCR) was used to amplify a reference 4 bp shorter than the original BCR ABL cDNA sequence. To achieve the purpose of site-directed mutagenesis. Enzymatic analysis showed that both types of BCR ABL mRNA can be used to obtain the corresponding cDNA reference. The co-amplification product of the reference and the template to be tested can be baseline separated by capillary electrophoresis, demonstrating its feasibility. This reference can be used for competitive PCR of BCR ABL fusion gene in chronic myelogenous leukemia, and this gene can be quantitatively detected