A matrix for DNA separation: Genotyping and sequencing using poly(vinylpyrrolidone) solution in uncoated capillaries
A matrix for DNA separation: Genotyping and sequencing using poly(vinylpyrrolidone) solution in uncoated capillaries
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DOI：
10.1021/ac970999h
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发表时间：
1998-04-01
影响因子：
7.4
通讯作者：
Yeung, ES
中科院分区：
文献类型：
--
作者：
Gao, QF;Yeung, ES
我们报告了一种新的筛选矩阵的基础上，商业化的聚（乙烯吡咯烷酮）的DNA分离。新的筛分基质在中等浓度下具有非常低的粘度，27 cP，其优异的自涂层性能可将电渗流降低到可忽略的水平，柱再生非常简单有效，在未涂层毛细管中实现了成功的分离。对于基因分型，我们表明D1 S80和釉原蛋白性别决定系统可以作为双链DNA进行基线分离，而vWF、TH 01、TPOX和CSF 1PO短串联重复序列可以作为单链DNA以单碱基分辨率分离在这个新矩阵中。M13mp18的测序显示出良好的分辨率高达500个碱基的高分子量馏分从市售PVP提取的溶液中。适应多毛细管阵列系统的可行性进行了讨论。
We report a new sieving matrix for DNA separation based on commercially available poly(vinylpyrrolidone). The new sieving matrix has a very low viscosity at moderate concentrations, e.g., 27 cP at 4.5%, Its excellent self-coating property can reduce electroosmotic flow to a negligible level, Column regeneration between runs is very simple and effective, Successful separations were achieved in uncoated capillaries. For genotyping, we show that D1S80 and amelogenin sex determination system can be baseline separated as double-stranded DNA, and vWF, TH01, TPOX, and CSF1PO short tandem repeats can be separated with single-base resolution as single-stranded DNA in this new matrix. Sequencing of M13mp18 showed good resolution up to 500 bases in a solution of high-molecular-weight fraction extracted from commercially available PVP. The feasibility of adaptation to a multicapillary array system is discussed.