Microarray-based DNA resequencing using 3' blocked primers.

Microarray-based DNA resequencing using 3' blocked primers.
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使用 3 封闭引物进行基于微阵列的 DNA 重测序。

DOI:
10.1016/j.ab.2007.10.044
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发表时间:
2008
影响因子:
2.9
通讯作者:
Liu,Qiang
Liu,Qiang
中科院分区:
生物学4区
文献类型:
--
作者:
Sram,Jakub;Sommer,SteveS;Liu,Qiang

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为了超越传统测序技术的通量和准确性,我们测试了一种使用3 '阻断引物(P∗s)的核酸扩增方法(焦磷裂解活化聚合[PAP])。作为原理证明,我们用P * s微阵列重新测序了因子IX基因的20bp区域。P * s对3′端错配具有超高特异性,对3′端长度的错配具有高特异性。我们正确地鉴定了两个野生型样本以及所有错配,包括三个单碱基替换,一个微缺失,一个微插入和一个杂合突变。尽管引物纯度有限,但匹配和不匹配的P * s之间的信噪比有时超过1000。因此,PAP重测序在基于微阵列的精确和高通量重测序方面显示出巨大的潜力。
To exceed the throughput and accuracy of conventional sequencing technologies, we tested a method (pyrophosphorolysis-activated polymerization [PAP]) of nucleic acid amplification that uses 3′ blocked primers (P∗s). As proof-of-principle, we resequenced a 20-bp region of the factor IX gene with a microarray of P∗s. P∗s discriminate 3′ end mismatches with ultra-high specificity as well as mismatches along their lengths with high specificity. We correctly identified two wild-type samples as well as all mismatches, including three single-base substitutions, one microdeletion, one microinsertion, and one heterozygous mutation. Despite limitations in the primer purity, the signal/noise ratio between the matched and mismatched P∗s sometimes exceeded 1000. Thus, PAP resequencing shows great potential for accurate and high-throughput microarray-based resequencing.
DOI: 10.1016/j.mrfmmm.2004.03.009
发表时间: 2004-10-04
影响因子: 2.3
作者:
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通讯作者: Lundeberg, J
DOI: 10.1093/nar/30.2.598
发表时间: 2002-01-15
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发表时间: 1979
影响因子: 14.9
作者:
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