Garbutt G J, Wilson J T, Schuster G S, Leary J J, Ward D C
Clin Chem. 1985 Jul;31(7):1203-6.
Earlier, we reported that the 5' end of the normal beta-globin gene (beta) resides on a 1.14-kilobase DNA fragment, whereas the 5' end of the sickle cell gene (beta s) resides on a 1.34-kilobase fragment. In that blot hybridization analysis, we used genomic DNA digested with restricted endonuclease Mst II, and radioactive probes with short half-life. We demonstrate here that, if a biotinylated probe is used instead in a slightly modified procedure, sickle cell anemia can be quickly and directly detected if there is as much as 5 micrograms of total genomic DNA in the sample. This procedure obviates the special precautions necessary when radioactive materials are used.
早些时候,我们报道正常β-珠蛋白基因(β)的5'端位于一个1.14千碱基的DNA片段上,而镰状细胞基因(βs)的5'端位于一个1.34千碱基的片段上。在那次印迹杂交分析中,我们使用了经限制性内切酶Mst II消化的基因组DNA以及半衰期短的放射性探针。我们在此证明,如果在一个稍有改进的程序中改用生物素化探针,那么样本中只要有多达5微克的总基因组DNA,就能快速直接检测出镰状细胞贫血。这个程序避免了使用放射性材料时所需的特殊预防措施。