Novel cyanine dye-labeled dideoxynucleoside triphosphates for DNA sequencing
Novel cyanine dye-labeled dideoxynucleoside triphosphates for DNA sequencing
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DOI：
10.1021/bc015522y
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发表时间：
2002-07-01
影响因子：
4.7
通讯作者：
Fuller, CW
中科院分区：
文献类型：
--
作者：
Duthie, RS;Kalve, IM;Fuller, CW
合成了单色菁染料标记的(Cy 5.0和Cy 5.5)双脱氧核苷-5‘-三磷酸，或’终止子‘，它们含有不同的间隔基长度，并在使用改进的热稳定性DNA聚合酶进行DNA测序方法中的有效性进行了评估。将单色菁染料终止子配制成两组独立的测序混合物，一组用于Cy 5.0，另一组用于Cy 5.5，并在不同的自动测序平台上进行评估。每组混合物包括两个具有17个原子连接子的嘧啶终止子和两个在染料和核苷酸之间具有10个原子连接子的嘌呤终止子。为这项循环测序研究选择的两组菁染料标记的终止子产生了改进的带型，其条带均一性类似于染料-引物法测序得到的带型。
Single color cyanine dye-labeled (Cy 5.0 and Cy 5.5) dideoxynucleoside-5'-triphosphates, or 'terminators', containing different spacer lengths were synthesized and evaluated for efficacy in DNA sequencing methods using a modified thermally stable DNA polymerase. The single color cyanine dye terminators were formulated into two separate sets of sequencing mixes, one for Cy 5.0 and the other for Cy 5.5, and evaluated on different automated sequencing platforms. Each set of mixes included two pyrimidine terminators with 17-atom linkers and two purine terminators with 10-atom linkers between the dye and the nucleotide. The two sets of cyanine dye-labeled terminators chosen for this cycle sequencing study produced improved band patterns with band uniformity similar to that obtained with dye-primer sequencing methods.