PEG-labeled nucleotides and nanopore detection for single molecule DNA sequencing by synthesis.
PEG-labeled nucleotides and nanopore detection for single molecule DNA sequencing by synthesis.
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DOI：
10.1038/srep00684
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发表时间：
2012
影响因子：
4.6
通讯作者：
Ju, Jingyue
中科院分区：
文献类型：
--
作者：
Kumar, Shiv;Tao, Chuanjuan;Chien, Minchen;Hellner, Brittney;Balijepalli, Arvind;Robertson, Joseph W. F.;Li, Zengmin;Russo, James J.;Reiner, Joseph E.;Kasianowicz, John J.;Ju, Jingyue
我们描述了一种新的基于单分子纳米孔的合成测序（Nano-SBS）策略，该策略可以通过检测从5′-磷酸修饰的核苷酸释放的4个不同大小的标签来准确区分4个碱基。基本原理如下。当每个核苷酸在聚合酶反应期间被掺入到生长的DNA链中时，其标签被释放并以释放顺序进入纳米孔。由于标签独特的化学结构，这产生了独特的离子电流阻断特征，从而以单碱基分辨率在单分子水平上以电子方式确定DNA序列。作为原理证明，我们将四种不同长度的PEG-香豆素标签连接到2′-脱氧鸟苷-5 ′-四磷酸的末端磷酸。我们证明了在聚合酶反应过程中核苷酸类似物的有效，准确的掺入，以及基于纳米孔离子电流的四个标签之间的出色区分。这种方法与以阵列形式连接到纳米孔的聚合酶结合，应该产生单分子电子Nano-SBS平台。
We describe a novel single molecule nanopore-based sequencing by synthesis (Nano-SBS) strategy that can accurately distinguish four bases by detecting 4 different sized tags released from 5′-phosphate-modified nucleotides. The basic principle is as follows. As each nucleotide is incorporated into the growing DNA strand during the polymerase reaction, its tag is released and enters a nanopore in release order. This produces a unique ionic current blockade signature due to the tag's distinct chemical structure, thereby determining DNA sequence electronically at single molecule level with single base resolution. As proof of principle, we attached four different length PEG-coumarin tags to the terminal phosphate of 2′-deoxyguanosine-5′-tetraphosphate. We demonstrate efficient, accurate incorporation of the nucleotide analogs during the polymerase reaction, and excellent discrimination among the four tags based on nanopore ionic currents. This approach coupled with polymerase attached to the nanopores in an array format should yield a single-molecule electronic Nano-SBS platform.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
38.3
作者：
通讯作者：
--
影响因子：
10.8
作者：
McNally B;Singer A;Yu Z;Sun Y;Weng Z;Meller A
通讯作者：
Meller A
影响因子：
8.6
作者：
Henrickson, SE;Misakian, M;Kasianowicz, JJ
通讯作者：
Kasianowicz, JJ
DOI：
10.1073/pnas.1001831107
发表时间：
2010-09-14
影响因子：
11.1
作者：
Derrington, Ian M.;Butler, Tom Z.;Gundlach, Jens H.
通讯作者：
Gundlach, Jens H.