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Kavya Kottapalli¹, Hsu Chao¹, Qi Jiang¹

¹Human Genome Sequencing Center, Baylor College of Medicine, Houston, TX, USA.

|November 26, 2025

概括

一种新的定量聚合酶链反应 (qPCR) 试验准确地预测了测序前RNA测序库中的核糖体RNA (rRNA) 含量. 这种具有成本效益的方法通过评估rRNA枯竭效率来确保可靠的转录组概况.

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:

核糖体RNA (rRNA) 构成了细胞RNA的主要部分,因此需要通过RNA测序 (RNA-Seq) 进行高效的去除,以便通过RNA测序 (RNA-Seq) 进行准确的转录组分析.
在RNA-Seq库的准备中,rRNA的低效耗尽可能会损害低丰度信使RNA (mRNA) 的精确测序.
目前用于评估rRNA内容预测序的方法通常不可靠,昂贵或缺乏可扩展性.

研究的目的:

在测序之前开发和验证可扩展,具有成本效益的测试方法,用于在RNA-Seq库中量化rRNA耗尽效率.
建立一种可靠的方法来预测在预测序列库中的rRNA读数的百分比.

主要方法:

开发一种实时定量聚合酶连锁反应 (qPCR) 试验,针对人类18S rRNA.
使用普遍人体参考 (UHR) 控制的连续稀释,优化qPCR效率.
使用来自644个图书馆的试点数据确定Ct值.
使用1748个人类总RNA-Seq和445个多A+RNA-Seq库进行验证.
使用来自四家供应商的Oligo (dT) 珠子评估mRNA丰富性能.

主要成果:

18S rRNA qPCR试验表明,在不同类型的RNA-Seq库中,序列化前的rRNA估计和序列化后的rRNA读取百分比之间存在强烈的相关性.
该试验在评估不同mRNA丰富方法的性能方面被证明是有效的.

结论:

开发的18S rRNA qPCR试验提供了一个可靠,可扩展和具有成本效益的解决方案,用于评估RNA-Seq库中的rRNA枯竭效率.
这种测试使研究人员能够在进行昂贵的测序之前预测和潜在地减轻与rRNA污染有关的问题.
该方法支持在转录组分析研究中提高准确性和效率.

关键词:

在18S qPCRR中.这就是Illumina Illumina.奥利戈 (dT) 珠子在 Poly A++ 中.在RNA-Seqq.总的RNA总量是多少耗尽效率的消耗效率是什么删除 rRNA 的过程.

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