Horvat S, Bünger L
Roslin Institute (Edinburgh), Division of Molecular Biology, UK.
Lab Anim. 1999 Oct;33(4):380-4. doi: 10.1258/002367799780487850.
A PCR-RFLP assay for genotyping at the mouse leptin receptor (Lepr(db)) mutation site was developed using modified primers. The first modified primer creates an AccI restriction site in the mutant Lepr(db) allele to distinguish between the Lepr(db) and Lepr+ alleles whereas the second modified primer creates another AccI site in both alleles to serve as a control for restriction enzyme digestion. The assay is robust and works efficiently on unpurified lysates of mouse tissues and can be applied at any age of the animal. The assay may be used as a diagnostic tool for maintenance of stocks, introgression or other types of crosses involving the Lepr(db) mutation.
利用改良引物开发了一种用于小鼠瘦素受体（Lepr(db)）突变位点基因分型的PCR-RFLP分析方法。第一个改良引物在突变的Lepr(db)等位基因中创建一个AccI限制位点，以区分Lepr(db)和Lepr+等位基因，而第二个改良引物在两个等位基因中都创建另一个AccI位点，用作限制酶消化的对照。该分析方法稳健，可在未纯化的小鼠组织裂解物上高效运行，并且可应用于动物的任何年龄。该分析方法可用作维持种群、基因渗入或涉及Lepr(db)突变的其他类型杂交的诊断工具。
