CN107002080B - 一种基于多重pcr的目标区域富集方法和试剂 - Google Patents
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Abstract
Description
组分 | 用量(μL) |
片段化DNA | X(100ng) |
T4DNA聚合酶(NEB公司) | 5 |
T4PNK(NEB公司) | 5 |
dNTP混合物 | 1.6 |
T4 PNK buffer(NEB公司)(10×) | 10 |
总量 | 100 |
组分 | 用量(μL) |
末端修复后DNA | X |
T4DNA连接酶(NEB公司) | 1 |
T4DNA连接酶缓冲液(NEB公司) | 5 |
第一接头 | 5 |
第二接头 | 5 |
ddH<sub>2</sub>O | 余量 |
总量 | 50 |
组分 | 用量(μL) |
捕获的单链DNA | 6 |
5×Ion AmpliSeq<sup>TM</sup> HiFi Mix(life technology) | 4 |
第三引物 | 10 |
总量 | 20 |
组分 | 用量(μL) |
线性扩增后的DNA | 6 |
5×Ion AmpliSeq<sup>TM</sup> HiFi Mix(life technology) | 4 |
第一引物和第三引物 | 10 |
总量 | 20 |
参数 | 数值 |
样本 | 外周血肿瘤细胞样本 |
总的读取数目 | 1253091 |
靶数目 | 1009600 |
靶数目比例 | 80.56% |
靶数目覆盖度 | 99.97% |
目标区域捕获效率 | 66% |
单碱基突变数目 | 43893 |
参数 | 数值 |
样本 | 肺癌样本 |
总的读取数目 | 3163794 |
平均测序深度 | 32204.13 |
靶数目覆盖度 | 100.00% |
目标区域捕获效率 | 89.10% |
Claims (10)
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CN201410796113 | 2014-12-18 | ||
CN2014107961137 | 2014-12-18 | ||
PCT/CN2015/096790 WO2016095736A1 (zh) | 2014-12-18 | 2015-12-09 | 一种基于多重pcr的目标区域富集方法和试剂 |
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CN107002080A CN107002080A (zh) | 2017-08-01 |
CN107002080B true CN107002080B (zh) | 2020-11-06 |
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US (1) | US10435736B2 (zh) |
CN (1) | CN107002080B (zh) |
WO (1) | WO2016095736A1 (zh) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
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CN108192955B (zh) * | 2018-01-17 | 2021-11-02 | 湖南大地同年生物科技有限公司 | 一种低频突变dna片段检测方法及文库建立方法 |
CN110240999B (zh) * | 2018-03-09 | 2022-09-06 | 浙江品级基因科技有限公司 | 一种提高循环肿瘤dna检出率的检测装置及方法 |
CN110241209B (zh) * | 2018-03-09 | 2022-11-29 | 浙江品级基因科技有限公司 | 一种引物、试剂盒及用途 |
WO2019205132A1 (zh) * | 2018-04-28 | 2019-10-31 | 深圳华大生命科学研究院 | 一种胎儿游离核酸的富集方法及其应用 |
CN110699426B (zh) * | 2019-01-02 | 2022-01-28 | 上海臻迪基因科技有限公司 | 基因目标区域富集方法及试剂盒 |
CN111041563B (zh) * | 2019-12-31 | 2023-07-21 | 广州精科医学检验所有限公司 | 一种靶向序列捕获及pcr建库方法 |
WO2021219114A1 (zh) * | 2020-04-30 | 2021-11-04 | 深圳市真迈生物科技有限公司 | 测序方法及其分析方法和***、计算机可读存储介质和电子设备 |
CN114015757B (zh) * | 2021-08-31 | 2024-02-02 | 江门市灿明生物科技有限公司 | 检测融合基因的方法 |
Citations (5)
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CN101815792A (zh) * | 2007-06-22 | 2010-08-25 | 纽约市哥伦比亚大学托管会 | 肿瘤特异性dna序列的特异性扩增 |
CN102329876A (zh) * | 2011-10-14 | 2012-01-25 | 深圳华大基因科技有限公司 | 一种测定待检测样本中疾病相关核酸分子的核苷酸序列的方法 |
CN102628082A (zh) * | 2012-04-10 | 2012-08-08 | 凯晶生物科技(苏州)有限公司 | 基于高通量测序技术进行核酸定性定量检测的方法 |
CN103572378A (zh) * | 2013-10-28 | 2014-02-12 | 广州爱健生物技术有限公司 | 基于Ion ProtonTM测序平台的小片段DNA文库的构建方法及其应用 |
CN104093890A (zh) * | 2012-01-26 | 2014-10-08 | 纽亘技术公司 | 用于靶向核酸序列富集和高效文库产生的组合物和方法 |
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DE19518505A1 (de) * | 1995-05-19 | 1996-11-21 | Max Planck Gesellschaft | Verfahren zur Genexpressionsanalyse |
CN101421410A (zh) | 2006-03-06 | 2009-04-29 | 纽约市哥伦比亚大学托管会 | 从混合的胎儿-母体来源中特异性扩增胎儿dna序列 |
CN102533985B (zh) | 2011-12-19 | 2014-08-06 | 深圳华大基因科技有限公司 | 一种检测dmd基因外显子缺失和/或重复的方法 |
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- 2015-12-09 US US15/537,396 patent/US10435736B2/en active Active
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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
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CN101815792A (zh) * | 2007-06-22 | 2010-08-25 | 纽约市哥伦比亚大学托管会 | 肿瘤特异性dna序列的特异性扩增 |
CN102329876A (zh) * | 2011-10-14 | 2012-01-25 | 深圳华大基因科技有限公司 | 一种测定待检测样本中疾病相关核酸分子的核苷酸序列的方法 |
CN104093890A (zh) * | 2012-01-26 | 2014-10-08 | 纽亘技术公司 | 用于靶向核酸序列富集和高效文库产生的组合物和方法 |
CN102628082A (zh) * | 2012-04-10 | 2012-08-08 | 凯晶生物科技(苏州)有限公司 | 基于高通量测序技术进行核酸定性定量检测的方法 |
CN103572378A (zh) * | 2013-10-28 | 2014-02-12 | 广州爱健生物技术有限公司 | 基于Ion ProtonTM测序平台的小片段DNA文库的构建方法及其应用 |
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WO2016095736A1 (zh) | 2016-06-23 |
US20180163251A1 (en) | 2018-06-14 |
CN107002080A (zh) | 2017-08-01 |
US10435736B2 (en) | 2019-10-08 |
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