A chemically synthesized peptoid-based drag-tag enhances free-solution DNA sequencing by capillary electrophoresis.
A chemically synthesized peptoid-based drag-tag enhances free-solution DNA sequencing by capillary electrophoresis.
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DOI：
10.1002/bip.21627
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发表时间：
2011
期刊：
影响因子：
2.9
通讯作者：
Barron, Annelise E.
中科院分区：
文献类型：
--
作者：
Haynes, Russell D.;Meagher, Robert J.;Barron, Annelise E.
We report a capillary-based DNA sequencing read length of 100 bases in 16 min using end-labeled free-solution conjugate electrophoresis (FSCE) with a monodisperse poly-N-substituted glycine (polypeptoid) as a synthetic drag-tag. FSCE enabled rapid separation of single-stranded (ss) DNA sequencing fragments with single-base resolution without the need for a viscous DNA separation matrix. Protein-based drag-tags previously used for FSCE sequencing, for example, streptavidin, are heterogeneous in molar mass (polydisperse); the resultant band-broadening can make it difficult to obtain the single-base resolution necessary for DNA sequencing. In this study, we synthesized and HPLC-purified a 70mer poly-N-(methoxyethyl)glycine (NMEG) drag-tag with a molar mass of~11 kDa. The NMEG monomers that comprise this peptoid drag-tag are interesting for bioanalytical applications, because the methoxyethyl side chain's chemical structure is reminiscent of the basic monomer unit of polyethylene glycol, a highly biocompatible commercially available polymer, which, however, is not available in monodisperse preparation at an ~11 kDa molar mass. This is the first report of ssDNA separation and of four-color, base-by-base DNA sequencing by FSCE through the use of a chemically synthesized drag-tag. These results show that high-molar mass, chemically synthesized drag-tags based on the polyNMEG structure, if obtained in monodisperse preparation, would serve as ideal drag-tags and could help FSCE reach the commercially relevant read lengths of 100 bases or more.
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影响因子：
3.2
作者：
Fowler SA;Blackwell HE
通讯作者：
Blackwell HE
影响因子：
2.9
作者：
Meagher, Robert J.;McCormick, Laurette C.;Barron, Annelise E.
通讯作者：
Barron, Annelise E.
DOI：
10.1073/pnas.89.20.9367
发表时间：
1992-10-15
影响因子：
11.1
作者：
SIMON, RJ;KANIA, RS;BARTLETT, PA
通讯作者：
BARTLETT, PA
影响因子：
5.5
作者：
Desruisseaux, C;Long, D;Slater, GW
通讯作者：
Slater, GW
影响因子：
1.8
作者：
Uno, T;Beausoleil, E;Zuckermann, RN
通讯作者：
Zuckermann, RN