用短读序列测序器和单管链接读取库方法对2-200千基基因组片段进行有针对性的分相处理
Veronika Mikhaylova1, Madison Rzepka1, Tetsuya Kawamura1
1Universal Sequencing Technology Corp., Carlsbad, CA, 92011, USA.
Scientific reports
|April 5, 2024
概括
使用TELL-Seq技术,可以使用短读序列测序,对目标基因组区域进行高达200kb的精确分相. 这种方法将DNA片段与分子条形码连接起来,方便对临床相关基因中异质合体位点的精确分析.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 生物信息学是一种生物信息学.
背景情况:
- 人类基因组中的异构位体代表了母体和父体染色体的等位体变异.
- 阶段化,或通过染色体拷贝解决等位基因差异,对于遗传分析至关重要.
- 现有的分相方法往往需要长距离的基因组信息,而短读测序器可能难以获得这些信息.
研究的目的:
- 为针对性基因组应用引入和验证TELL-Seq协议.
- 为了能够精确地分阶段的丰富的位置介于2到200千基 (kb).
- 证明TELL-Seq用于分析临床相关基因的实用性.
主要方法:
- 使用分子标识符 (条形码) 来生成从长DNA片段 (高达200kb) 中链接读取的TELL-Seq库编制.
- 使用CRISPR/Cas9介导切除,使用CRISPR/Cas9介导对外核酶消化进行保护,以及使用长期PCR,对特定位点进行向丰富.
- 在短读序列器上使用TELL-Seq协议对丰富的2-200 kb loci进行分阶段.
主要成果:
- 开发的TELL-Seq协议精确地分阶段将目标位点跨越了各种大小 (2-200 kb) 的范围.
- 通过使用各种方法,包括CRISPR/Cas9和长期PCR,丰富基因的分期成功实现了.
- 该协议在临床相关基因 (例如BRCA1,BRCA2,MLH1) 的目标区域内解决异构位的准确性很高.
结论:
- TELL-Seq是一种使用短读序列的多功能和准确的基因组分相定向方法.
- 该协议有助于解决临床重要基因中复杂的基因组变异.
- 这种方法增强了短读测序的能力,用于全面的遗传分析.
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