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使用二色和四色基下一代测序平台检测同聚合物的性能

HuiJuan Chen^1,2,3, Bing Wang¹, LiLi Cai¹

¹Beijing ChosenMed Clinical Laboratory Co. Ltd, Beijing, 100176, China.

|May 31, 2024

概括

下一代测序 (NGS) 平台在同聚合物 (HP) 测序精度方面扎. 独特的分子标识符 (UMIs) 在多个NGS系统中显著提高HP测序性能,提高数据可靠性.

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:

同聚合物 (HP) 区域对下一代测序 (NGS) 具有挑战性,因为其错误率很高,包括插入,删除和替换.
评估不同NGS平台在精确测序这些HP区域方面的性能对于可靠的基因组分析至关重要.

研究的目的:

评估二色和四色化NGS平台在同聚合物区域测序中的性能.
在HP测序中比较MGISEQ-2000,MGISEQ-200和NextSeq 2000平台的准确性.
研究独特分子标识符 (UMI) 对提高HP测序精度的影响.

主要方法:

含有同聚合物区域的等离子体的构造.
连续稀释等离子体以产生具有不同HP频率 (3%,10%,30%,60%) 的样本.
在MGISEQ-2000,MGISEQ-200和NextSeq 2000平台上对这些样品进行测序,有或没有独特分子标识符 (UMI) 管道.

主要成果:

在所有测试平台上,在HP长度和检测频率之间观察到负相关性.
在所有系统中,所有8mer HP都显示了显著下降的检测率,除了3%频率的NextSeq 2000.
应用UMI在很大程度上将检测到的频率正常化到预期值,MGI 200平台上的poly-G则是少数例外,这表明性能有所改善.

结论:

MGISEQ-2000和NextSeq 2000平台在对同聚合物区域进行测序方面表现相似.
引入独特的分子标识符 (UMI) 是一种高度有效的策略,可以提高NGS平台在测序同聚合物区域的准确性.
这项研究为容易出现序列错误的区域优化NGS工作流提供了有价值的见解.

关键词:

同聚合物检测检测同聚合物检测这是下一代测序.测序平台的测序平台.

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