无胶毛细管电泳分离碱基对范围宽的DNA片段

SEPARATION OF WIDE RANGE DNA FRAGMENTS USING NON GEL CAPILLARY ELECTROPHORESIS

提出一种无胶毛细管电泳分离碱基对范围宽的ＤＮＡ片段的方法．用ＤＢ－１气相色谱毛细管柱，以羟乙基纤维素为筛分介质，研究了λＤＮＡ／ＥｃｏＲⅠ＋ＨｉｎｄⅢ限制性片段的分离，分离效率达１．２×１０６板／ｍ，检测限为２．１×１０－１７ｍｏｌ．应用于一种鲤鱼种族鉴别基因的聚合酶链反应（ＰＣＲ）扩增产物的分离鉴定，结果与实际相符，分离速度比传统电泳方法提高２０倍．

This paper reported the separation of wide range DNA fragments using non gel capillary electrophoresis. In a DB 1 GC capillary ( 40 cm×100 μm,i.d.,effective length 33 cm ), using hydroxyethylcellulose (HEC) as sieving media, separation of λDNA/EcoRⅠ+HindⅢ fragments was studied. Efficiency of 1.2×10 6 (the number of theoretical plate per meter) was achieved, and the detection limit was 2.1×10 -17 mol. The polymerase chain reaction (PCR) product from a carp gene was detected, and the bp number of DNA fragments was determined. The results were consistent with the real length,but the analysis speed was 20 times higher than traditional electrophoresis method.