间隔子修饰引物对扩增产物电泳行为的作用初探  

Preliminary Study of the Action on Electrophoresis by DNA Primer with Spacer Modification

作　　者：孙敬[1] 赵兴春[1] 叶健[1] 胡兰[1] 李彩霞[1] 欧元[1] 赵蕾[1] 王乐[1] 刘琳[1] 王雪倩[1] 赵雯雯[1] 

机构地区：[1]公安部物证鉴定中心,北京100038

出　　处：《生物技术通讯》2015年第1期107-110,共4页Letters in Biotechnology

基　　金：公安部科技强警基础工作专项(2012GABJC032);基本科研业务费专项(2012JBYY006)

摘　　要：目的:初步研究利用C3Spacer间隔子修饰引物对扩增产物电泳行为的影响及作用。方法:选取1个常用短串联重复序列(STR)位点,利用间隔子修饰该位点荧光标记引物,以DNA标准物质为模板进行PCR扩增,记录相应扩增产物DNA片段长度,进行修饰与长度变化的相关性分析。结果:选取STR位点D13S317,分别利用TTTTC3SpacerC3Spacer、TTTTC3Spacer、TTTT修饰R0X标记引物,相应扩增产物长度为182.67±0.05、182.19±0.11和181.6±0.19bp,未进行修饰的对照组引物扩增产物长度为177.09±0.15 bp,产物DNA片段长度随不同修饰基团的修饰发生规律性变化。结论:发现了一种修饰基团,用该基团修饰引物后,可在体外通过PCR反应改变扩增产物等位基因片段的大小,从而在不改变特异性引物信息的前提下使产物发生规律性位移,修饰基团与DNA片段大小呈内在相关性。Objective： To develop the preliminary study of the action on electrophoresis by DNA primer with spacer modification. Methods： One STR locus was selected. The primers labeled by ROX was modified. PCR was performed using 9947A as DNA template. DNA length of PCR products was calculated. The relationship between modification and DNA length was analyzed. Results： D13S317 primers labeled by ROX was modified respectively with TTTTC3SpacerC3Spacer, TTTTC3Spacer and TTTT, and the corresponding DNA length was 182.67±0.05, 182.19±0.11 and 181.6±0.19 bp. The control DNA length without modification was 177.09±0.15 bp. The results indicated that the DNA length of PCR products varied with different modification groups. Conelusion： One kind of primer modification that can affect the DNA length of PCR products was discovered without the alteration of specific primer length and sequences.

关 键 词：间隔子 修饰 引物 电泳 

分 类 号：Q78[生物学—分子生物学]