A single molecule array for digital targeted molecular analyses.
A single molecule array for digital targeted molecular analyses.
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DOI：
10.1093/nar/gkn921
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发表时间：
2009-01
影响因子：
14.9
通讯作者：
Nilsson M
中科院分区：
文献类型：
--
作者：
Göransson J;Wählby C;Isaksson M;Howell WM;Jarvius J;Nilsson M
我们提出了一种新的随机数组格式和解码方案，用于目标多路数字分子分析。DNA样本分析使用多套挂锁或选择探针，在目标识别时产生环状DNA分子。环状DNA分子通过滚环扩增（RCA）得到扩增单分子（asm）。将所有asm固定在显微镜载玻片上，产生随机阵列。根据组合解码方案，通过小组标记探针的连续杂交来识别和计数asm。我们表明，随机阵列格式允许杂交、成像和去杂交至少10次迭代，这是组合解码方案所需的一个过程。我们进一步研究了随机数组格式的定量动态范围和精度。最后，作为示范，将该解码方案应用于拷贝数变异已验证的样本中基因组位点的多重定量分析。在所分析的31个基因座中，除1个外，其余均被正确识别，并根据已知的拷贝数变化做出反应。解码策略是通用的，因为目标可以是任何通过分子探测反应编码成DNA环的生物分子。
We present a new random array format together with a decoding scheme for targeted multiplex digital molecular analyses. DNA samples are analyzed using multiplex sets of padlock or selector probes that create circular DNA molecules upon target recognition. The circularized DNA molecules are amplified through rolling-circle amplification (RCA) to generate amplified single molecules (ASMs). A random array is generated by immobilizing all ASMs on a microscopy glass slide. The ASMs are identified and counted through serial hybridizations of small sets of tag probes, according to a combinatorial decoding scheme. We show that random array format permits at least 10 iterations of hybridization, imaging and dehybridization, a process required for the combinatorial decoding scheme. We further investigated the quantitative dynamic range and precision of the random array format. Finally, as a demonstration, the decoding scheme was applied for multiplex quantitative analysis of genomic loci in samples having verified copy-number variations. Of 31 analyzed loci, all but one were correctly identified and responded according to the known copy-number variations. The decoding strategy is generic in that the target can be any biomolecule which has been encoded into a DNA circle via a molecular probing reaction.
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影响因子：
9.3
作者：
Fredriksson, Simon;Horecka, Joe;Davis, Ronald W.
通讯作者：
Davis, Ronald W.
DOI：
10.1073/pnas.0702165104
发表时间：
2007-05-29
影响因子：
11.1
作者：
Dahl, Fredrik;Stenberg, Johan;Ji, Hanlee
通讯作者：
Ji, Hanlee
影响因子：
14.9
作者：
Isaksson M;Stenberg J;Dahl F;Thuresson AC;Bondeson ML;Nilsson M
通讯作者：
Nilsson M
影响因子：
46.9
作者：
Fredriksson, S;Gullberg, M;Landegren, U
通讯作者：
Landegren, U
影响因子：
14.9
作者：
CHETVERINA, HV;CHETVERIN, AB
通讯作者：
CHETVERIN, AB