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本文描述用一种标准限制核酸内切酶技术的改良法,检测镰状突变基因以快速诊断风险妊娠的镰状细胞贫血。方法是先从周围血自细胞或羊膜细胞制备 DNA,含镰状细胞突变基因(β~6GAG→GTG)的DNA 靶顺序在引物介导下,由 DNA 多聚酶Ⅰ催化扩增20万倍,为鉴别正常β~6GAG 与病理 GTG,先对扩增了的 DNA 作快速液相杂交,探针是能与β~A 非编码链互补、5′末端~(32)P 标记的40硷基片段,为了分析β~6编码的另一种点突变——导致 Hbc 的GAG→AAG 突变,还使用了β~c 特异性40硷基杂
This article describes a sickle cell anemia test that detects sickle mutant genes for rapid diagnosis of at-risk pregnancies using a modified version of a standard restriction endonuclease technique. The method is to prepare DNA from cells or amnion cells from peripheral blood. The DNA target sequence containing the mutant gene of sickle cell (β6GAG → GTG) is 200K times amplified by DNA polymerase Ⅰ The normal β ~ 6GAG and pathological GTG were firstly rapidly hybridized to the amplified DNA. The probe was a 40-base fragment complementary to the non-coding β ~ A chain and labeled with ~ (32) P at the 5 ’end. Analysis of another point mutation in the beta 6 coding - resulted in a GAG → AAG mutation of Hbc, as well as a beta-c specific 40-base-miscellaneous