全球全基因组牛津纳米孔测序SARS-CoV-2使用式安普利康
Ruslan Kalendar1,2, Ulykbek Kairov3, Daniyar Karabayev3,4
1Center for Life Sciences, National Laboratory Astana, Nazarbayev University, Astana, Kazakhstan. ruslan.kalendar@helsinki.fi.
Scientific reports
|June 26, 2023
概括
我们在牛津纳米孔平台上开发了一套通用原始工具,用于在牛津纳米孔平台上快速测序SARS-CoV-2基因组. 这种优化的协议使得成本效益高的全基因组测序,即使从退化的RNA样本.
科学领域:
- 基因组学就是基因组学.
- 分子生物学分子生物学
- 病毒学 病毒学
背景情况:
- 准确和高效的SARS-CoV-2基因组测序对于流行病学监测和变种追踪至关重要.
- 现有的SARS-CoV-2基因组测序方法可能耗时且昂贵,特别是在低滴度或退化样本.
研究的目的:
- 使用牛津纳米孔技术开发一个全面的,多重复合的原始套件,用于使用牛津纳米孔技术进行通用SARS-CoV-2基因组测序.
- 优化cDNA合成协议,以获得高产量和长度cDNA的多种RNA质量.
- 为现有测序方法提供更快,更具成本效益的替代方案.
主要方法:
- 开发一个可与牛津纳米孔快速条形码库套件兼容的多重化原料套件.
- 使用Maxima H减去逆转录酶和SARS-CoV-2特定原料的cDNA合成协议的优化.
- 单片或双片安普利康 (1.24.8 kb) 的应用,用于全基因组和向测序.
主要成果:
- 开发的原始套件使得通用SARS-CoV-2基因组测序与牛津纳米孔.
- 优化的协议产生了大量的cDNA,促进了低滴度和退化RNA样本的测序.
- 该协议支持全基因组测序,使用最长可达4.8kb的amplicon.
结论:
- 新的基础套件和优化的协议为SARS-CoV-2基因组测序提供了一种高效和成本效益的方法.
- 与午夜多重PCR方法相比,这种方法可以减少周转时间和成本.
- 该协议具有多功能性,适用于SARS-CoV-2的全基因组和向测序.
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