Extraction and processing of circulating DNA from large sample volumes using methylation on beads for the detection of rare epigenetic events.
Extraction and processing of circulating DNA from large sample volumes using methylation on beads for the detection of rare epigenetic events.
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DOI：
10.1016/j.cca.2013.07.023
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发表时间：
2013-10-21
期刊：
影响因子：
--
通讯作者：
Wang TH
中科院分区：
文献类型：
--
作者：
Keeley B;Stark A;Pisanic TR 2nd;Kwak R;Zhang Y;Wrangle J;Baylin S;Herman J;Ahuja N;Brock MV;Wang TH
甲基化的肿瘤特异性循环DNA作为潜在的癌症生物标志物，但肿瘤特异性的循环DNA的相对稀缺性是对传统的DNA提取和加工技术的挑战。与传统相比包括苯酚氯仿和酒精提取的技术，珠子上的甲基化在提取效率上提高了1.5至5倍，导致PCR抑制剂的含量较小，从而使分析敏感性提高了25倍以上的敏感性。
The use of methylated tumor-specific circulating DNA has shown great promise as a potential cancer biomarker. Nonetheless, the relative scarcity of tumor-specific circulating DNA presents a challenge for traditional DNA extraction and processing techniques. Here we demonstrate a single tube extraction and processing technique dubbed “methylation on beads” that allows for DNA extraction and bisulfite conversion for up to 2 ml of plasma or serum. In comparison to traditional techniques including phenol chloroform and alcohol extraction, methylation on beads yields a 1.5 to 5-fold improvement in extraction efficiency. The technique results in far less carryover of PCR inhibitors yielding analytical sensitivity improvements of over 25-fold. The combination of improved recovery and sensitivity make possible the detection of rare epigenetic events and the development of high sensitivity epigenetic diagnostic assays.
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DOI：
10.1196/annals.1318.006
发表时间：
2004-01-01
期刊：
CIRCULATING NUCLEIC ACIDS IN PLASMA/SERUM III AND SERUM PROTEOMICS
影响因子：
--
作者：
Herman, JG
通讯作者：
Herman, JG
负载磁铁矿的二氧化硅微球的制备用于固相提取大豆食品中的基因组 DNA
影响因子：
4.1
作者：
Shi, Ruobing;Wang, Yucong;Wan, Qian-Hong
通讯作者：
Wan, Qian-Hong
DOI：
10.1073/pnas.0507904102
发表时间：
2005-11-08
影响因子：
11.1
作者：
Diehl, F;Li, M;Vogelstein, B
通讯作者：
Vogelstein, B
影响因子：
--
作者：
Hesson LB;Cooper WN;Latif F
通讯作者：
Latif F
影响因子：
158.5
作者：
Brock, Malcolm V.;Hooker, Craig M.;Herman, James G.
通讯作者：
Herman, James G.