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使用纳米孔直接的寡核酸测序
Sachin Chalapati1, Conor A Crosbie1, Dixita Limbachiya1
1Helixworks Technologies, Environmental Research Institute, University College Cork, Cork, T23 XE10, Ireland.
Open research Europe
|August 30, 2023
概括
研究人员使用MinION平台开发了一种用于单链DNA寡核酸的直接测序方法. 这种方法绕过了放大和第二链合成,使得新的DNA数据存储应用成为可能.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物技术是生物技术.
背景情况:
- 第三代DNA测序提供了核酸的直接分析.
- 单链DNA (ssDNA) 或RNA的测序提供了独特的见解,但面临着适配器兼容性挑战.
- 牛津纳米孔MiniON平台测序单一DNA链,但通常需要双链DNA (dsDNA) 库的准备.
研究的目的:
- 使用MinION平台开发一个用于短,单链寡核酸的直接测序方法.
- 为了实现无需放大或第二链合成的测序.
- 探索DNA数据存储中的应用.
主要方法:
- 一种新的图书馆制备方法,涉及5'化寡核酸的单个回火步骤.
- 设计适配器序列以结合寡核酸的5'端,创建一个3'腺突起.
- 利用MinION平台对准备好的寡核酸进行直接的纳米孔测序.
主要成果:
- 在没有放大或第二链合成的情况下,成功地直接测序短,单链的寡核酸.
- 从42,000个序列池中,大约90%的120nT直角序列得到了高准确度的恢复.
- 在纳米孔原始数据中识别潜在的工件,包括空信号和融合序列.
结论:
- 这种直接的寡核酸测序方法与基于结合的图书馆准备纳米孔测序是兼容的.
- 该技术克服了对单链核酸的适配器兼容性问题.
- 允许新型应用,特别是在使用寡核酸作为信息载体的DNA数据存储系统中.
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