DNA sequencing by delayed extraction matrix-assisted laser desorption/ionization time of flight mass spectrometry
DNA sequencing by delayed extraction matrix-assisted laser desorption/ionization time of flight mass spectrometry
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DOI：
10.1073/pnas.93.10.4724
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发表时间：
1996-05-14
影响因子：
11.1
通讯作者：
Haff, LA
中科院分区：
文献类型：
--
作者：
Roskey, MT;Juhasz, P;Haff, LA
基质辅助激光解吸/电离（MALDI）飞行时间质谱用于检测和排序由桑格双脱氧循环测序产生的DNA片段。这是通过使用延迟离子提取技术（DE-MALDI）提高MALDI方法的灵敏度和分辨率来实现的。优化循环测序化学以产生多达100 fmol的每个特异性双脱氧终止片段，其由在40-和50-碱基模板上退火的13-碱基引物的延伸产生。通过DE-MALDI对所得测序混合物的分析鉴定了适当的终止产物。该技术提供了一种新的非基于凝胶的DNA测序方法，与传统方法相比，该方法最终可能具有相当大的速度优势。
Matrix-assisted laser desorption/ionization (MALDI) time of flight mass spectrometry was used to detect and order DNA fragments generated by Sanger dideoxy cycle sequencing. This was accomplished by improving the sensitivity and resolution of the MALDI method using a delayed ion extraction technique (DE-MALDI). The cycle sequencing chemistry was optimized to produce as much as 100 fmol of each specific dideoxy terminated fragment, generated from extension of a 13-base primer annealed on 40- and 50-base templates, Analysis of the resultant sequencing mixture by DE-MALDI identified the appropriate termination products. The technique provides a new non-gel-based method to sequence DNA which may ultimately have considerable speed advantages over traditional methodologies.