Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen as fluorescence inducing agent.
Fast separation of single-stranded oligonucleotides by capillary electrophoresis using OliGreen as fluorescence inducing agent.
复制标题
使用 OliGreen 作为荧光诱导剂，通过毛细管电泳快速分离单链寡核苷酸。
DOI：
10.1002/elps.200500099
复制
发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Chu,Benjamin
中科院分区：
文献类型：
--
作者：
Zhang,Jun;Liang,Dehai;He,Weidong;Wan,Fen;Ying,Qicong;Chu,Benjamin
介绍了用OliGreen®进行CE快速分离寡核苷酸（oligos）大小标记物的方法，包括分离介质和尿素变性剂浓度的影响。对低聚物的分离，发现低聚绿染料比溴化乙啶更敏感（基于S/N考虑，灵敏度约为6倍）。当F127在1×Tris -硼酸- EDTA （TBE）中浓度达到30% w/v时，低聚物分离的分辨率更高。在分离介质中加入尿素会降低灵敏度。在优化的运行条件下，以30% w/v F127/1×TBE溶液为分离介质，在800 v /cm的电场下，在3 cm长的毛细管中，在1.3 min内以1碱基的分辨率成功分离寡糖大小标记物，这是迄今为止报道的在类似尺寸范围内最快的高分辨率毛细管凝胶电泳分离。
The fast separation of oligonucleotide (oligos) sizing marker by CE using OliGreen® and including effects due to the concentration of separation medium and urea denaturant is presented. OliGreen dye is found to be more sensitive than ethidium bromide (by a factor of about 6 based on S/N considerations) for the oligos' separations. Higher concentration of F127 in 1×Tris‐boricacid‐EDTA (TBE) up to 30% w/v leads to better resolution of oligos separations. The addition of urea into the separation medium decreases the sensitivity. With an optimized running condition, the oligos sizing marker could be successfully separated with 1‐base resolution within 1.3 min by using 30% w/v F127/1×TBE solution as the separation medium at an applied electric field of 800 V/cm in a 3 cm long capillary, the fastest capillary gel electrophoresis separation with high resolution reported to date for oligos in the similar size range.