在4.5厘米的塑料电泳微通道上进行高分辨率单链DNA分析。

High-resolution single-stranded DNA analysis on 4.5 cm plastic electrophoretic microchannels.

作者信息

Shi Yining, Anderson Rolfe C

机构信息

ACLARA BioSciences, Inc., Mountain View, CA 94043, USA.

出版信息

Electrophoresis. 2003 Oct;24(19-20):3371-7. doi: 10.1002/elps.200305553.

DOI:10.1002/elps.200305553

PMID:14595684

Abstract

Plastic microchannels (4.5 cm long) fabricated from an etched glass master were tested for high-resolution single-stranded DNA analysis. Using replaceable denaturing linear polyacrylamide as sieving matrix, one-color separation of a fragment sizing standard with single-base resolution (R > 0.5) was achieved up to 275 bases. Two-color sizing analysis of four loci short tandem repeat (STR) allelic ladder (CSF1PO, TPOX, TH01, vWA) with single-base resolution (R = 0.62) on TH01 alleles 9.3 (198 bp) and 10 (199 bp) was demonstrated. An average standard deviation of +/- 0.06 bp and +/-0.11 bp in sizing 32 alleles of the CTTv ladder was attained between runs and between channels, respectively. Four-color sequencing separation of a terminator sequencing standard showed a base-calling accuracy of 99.1% out to 320 bases in 13 min.

摘要

对由蚀刻玻璃母版制成的塑料微通道（4.5厘米长）进行了高分辨率单链DNA分析测试。使用可替换的变性线性聚丙烯酰胺作为筛分基质，在高达275个碱基的范围内实现了单碱基分辨率（R>0.5）的片段大小标准品的单色分离。展示了对四个位点短串联重复序列（STR）等位基因阶梯（CSF1PO、TPOX、TH01、vWA）进行双色调大小分析，在TH01等位基因9.3（198 bp）和10（199 bp）上具有单碱基分辨率（R = 0.62）。在CTTv阶梯的32个等位基因大小测定中，每次运行之间和通道之间的平均标准偏差分别为±0.06 bp和±0.11 bp。对终止子测序标准品进行的四色测序分离在13分钟内对320个碱基的碱基识别准确率达到99.1%。