Separation of long DNA fragments by capillary gel electrophoresis with laser‐induced fluorescence detection
Separation of long DNA fragments by capillary gel electrophoresis with laser‐induced fluorescence detection
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通过毛细管凝胶电泳和激光诱导荧光检测分离长 DNA 片段
DOI：
10.1002/elps.1150150136
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
H. Kambara
中科院分区：
文献类型：
--
作者：
T. Nishikawa;H. Kambara
通过毛细管凝胶电泳结合激光诱导荧光检测分离长DNA片段，并检查了获得DNA片段高分辨率分离的两种方法。一种方法以牺牲检测时间为代价实现了高分辨率，而另一种方法同时实现了高分辨率和快速检测。前一种方法使用300 cm长的毛细管，凝胶浓度为4%T，电场强度为70 V/cm。该电泳的分离极限为800个碱基，相邻峰的分离度为0.5。在这些条件下，测量时间为44 h。后一种方法使用200 cm长的毛细管，凝胶浓度为3% T，电场强度为170 V/cm。该电泳的分辨极限为680个碱基，相邻峰的分辨值为0.5，测量时间仅为10 h。使用这两种方法，具有小于600个碱基的片段被有效地分离。碱基数小于500的相邻峰的分离度值大于1.0，碱基数小于100的相邻峰的分离度值大于2.7。因此，这些方法提高了碱基序列测定的准确性。
Long DNA fragments were separated by capillary gel electrophoresis with laser‐induced fluorescence detection, and two approaches to obtaining high‐resolution separation of DNA fragments were examined. One method achieved high resolution at the expense of detection time, while the other achieved both high resolution and fast detection. The former approach used a 300 cm long capillary with a gel concentration of 4% T and an electric field strength of 70 V/cm. The resolution limit in this electrophoresis was 800 bases with a resolution value of 0.5 for adjacent peaks. Under these conditions, the measurement time was 44 h. The latter approach used a 200 cm long capillary with a gel concentration of 3% T and an electric field strength of 170 V/cm. The resolution limit in this electrophoresis was 680 bases with a resolution value of 0.5 for adjacent peaks, and the measurement time was only 10 h. With both approaches, fragments with less than 600 bases were efficiently separated. The resolution values for adjacent peaks with less than 500 bases are greater than 1.0, and those for peaks with less than 100 bases are greater than 2.7. These approaches thus improve the accuracy of the base sequence determination.
使用改良噬菌体 T7 DNA 聚合酶进行 DNA 序列分析。
DOI：
--
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Tabor,S;Richardson,CC
通讯作者：
Richardson,CC