原位PCR标本的处理及反应条件的优化  

Disposal of Specimen and the Optimization of Response Condition in PCR in Situ

作　　者：轩小燕[1] 李敏[1] 杜献堂[1] 

机构地区：[1]郑州大学医学院微生物学与免疫学教研室,河南郑州450052

出　　处：《河南职工医学院学报》2004年第3期218-219,共2页Journal of Henan Medical College For Staff and Workers

基　　金：国家自然科学基金资助项目 3 9970 119

摘　　要：目的　探讨原位PCR中标本处理的最佳方法和反应的最佳条件。方法　应用原位PCR方法对昆明小鼠进行基因检测和定位 ,对标本使用不同的方法处理 ,检测和定位过程中改变主要的反应条件。结果　进行原位PCR的标本使用 0 0 5 %多聚赖氨酸处理黏附性最好 ;4 %多聚甲醛固定标本 5min、30 μg/ml蛋白酶K 37℃处理 30min标本的结构和通透性最好 ;PCR反应液中Mg2 + 达到 4 5mmol/L扩增结果最好。Objective To explore the disposal of specimen and the optimization of response condition in PCR in situ.Methods Genes were detected and orientated in Kunming mice by PCR in situ.Results The adhesion of specimen was best by disposing of 0.05% poly-L-lysine; the structure and permeability were best by disposing of 4% paraformal-dehyde for 5minutes and by disposing of 30 μg/ml proteinase K 37℃ for 30 minutes; the better thickness of Mg 2+ was 4.5 mmol/L in PCR.Conclusion The structure and permeability of specimen and the response condition are important in PCR in situ.

关 键 词：原位PCR 标本处理 反应条件 条件优化 基因检测 

分 类 号：R394[医药卫生—医学遗传学]