CN107002080A - 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 |
T4PNK 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) |
DNA连接产物 | X |
dNTP(25mM) | 3 |
Platinum pfx DNA polymerase(life technology) | 0.6 |
第一引物 | 3 |
第二引物 | 3 |
MgSO4 | 4 |
10×pfx buffer | 10 |
ddH<sub>2</sub>O | 余量 |
总量 | 100 |
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)
- 一种基于多重PCR的目标区域富集方法,包括如下步骤:在连接酶作用下,在含待富集目标区域的核酸片段两端分别连接第一接头和第二接头,得到接头连接产物,其中所述第一接头包括用于后续PCR扩增的第一接头固定序列、用于标记不同样本的第一标签序列和用于标记不同目标区域分子序列的第二标签序列;使用特异性结合所述第一接头固定序列的第一引物和特异性结合所述第二接头的第二引物,对所述接头连接产物进行PCR扩增,得到扩增产物,其中所述第一引物或第二引物具有第一亲和标记;通过固相载体捕获所述扩增产物中带有所述第一亲和标记的单链,其中所述固相载体上带有能够与所述第一亲和标记亲和结合的第二亲和标记;以捕获的所述单链为模板,使用第三引物进行单引物线性扩增,其中所述第三引物包括5’端的第三引物固定序列和3’端的目标区域特异性结合序列;以所述线性扩增产物为模板,使用所述第三引物和所述第一引物进行指数扩增,得到包含目标区域的产物。
- 根据权利要求1所述的基于多重PCR的目标区域富集方法,其特征在于,所述第一标签序列为5-10个碱基长度的序列,所述第二标签序列为10-12个碱基长度的随机序列。
- 根据权利要求1所述的基于多重PCR的目标区域富集方法,其特征在于,所述含待富集目标区域的核酸片段为末端修复过的核酸片段。
- 根据权利要求1所述的基于多重PCR的目标区域富集方法,其特征在于,所述第一引物具有第一亲和标记。
- 根据权利要求1所述的基于多重PCR的目标区域富集方法,其特征在于,所述第一亲和标记为生物素标记,所述第二亲和标记为链霉亲和素标记,所述固相载体为磁珠。
- 一种基于多重PCR的目标区域富集试剂,包括如下组成部分:第一接头和第二接头,用于在连接酶作用下分别连接到含待富集目标区域的核酸片段两端,以得到接头连接产物,其中所述第一接头包括用于后续PCR扩增的第一接头固定序列、用于标记不同样本的第一标签序列和用于标记不同目标区域分子序列的第二标签序列;第一引物和第二引物,用于对所述接头连接产物进行PCR扩增,得到扩增 产物,其中所述第一引物特异性结合所述第一接头固定序列,所述第二引物特异性结合所述第二接头,并且所述第一引物或第二引物具有第一亲和标记;固相载体,用于捕获所述扩增产物中带有所述第一亲和标记的单链,其中所述固相载体上带有能够与所述第一亲和标记亲和结合的第二亲和标记;第三引物,用于以捕获的所述单链为模板,进行单引物线性扩增,其中所述第三引物包括5’端的第三引物固定序列和3’端的目标区域特异性结合序列;其中,所述第三引物和所述第一引物还用于以所述线性扩增产物为模板,进行指数扩增,得到包含目标区域的产物。
- 根据权利要求6所述的基于多重PCR的目标区域富集试剂,其特征在于,所述第一标签序列为5-10个碱基长度的序列,所述第二标签序列为10-12个碱基长度的随机序列。
- 根据权利要求6所述的基于多重PCR的目标区域富集试剂,其特征在于,所述含待富集目标区域的核酸片段为末端修复过的核酸片段。
- 根据权利要求6所述的基于多重PCR的目标区域富集试剂,其特征在于,所述第一引物具有第一亲和标记。
- 根据权利要求6所述的基于多重PCR的目标区域富集试剂,其特征在于,所述第一亲和标记为生物素标记,所述第二亲和标记为链霉亲和素标记,所述固相载体为磁珠。
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Cited By (4)
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CN108192955A (zh) * | 2018-01-17 | 2018-06-22 | 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 |
CN110241209A (zh) * | 2018-03-09 | 2019-09-17 | 林云富 | 一种引物、试剂盒及用途 |
CN110240999A (zh) * | 2018-03-09 | 2019-09-17 | 林云富 | 一种提高循环肿瘤dna检出率的检测装置及方法 |
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 |
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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 |
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE19518505A1 (de) * | 1995-05-19 | 1996-11-21 | Max Planck Gesellschaft | Verfahren zur Genexpressionsanalyse |
BRPI0709545A2 (pt) | 2006-03-06 | 2011-07-19 | Univ Columbia | amplificação especìfica de seqüência de dna fetal de uma mistura de origem fetal-maternal |
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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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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 |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108192955A (zh) * | 2018-01-17 | 2018-06-22 | 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 |
CN110241209A (zh) * | 2018-03-09 | 2019-09-17 | 林云富 | 一种引物、试剂盒及用途 |
CN110240999A (zh) * | 2018-03-09 | 2019-09-17 | 林云富 | 一种提高循环肿瘤dna检出率的检测装置及方法 |
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 |
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