通过快速的PRO-seq对新生RNA的分析来增强转录组映射
bioRxiv : the preprint server for biology
|May 20, 2024
概括
快速的PRO-seq (rPRO-seq) 简化了新生的转录分析,只需12小时就能够准确地绘制转录组图. 这种更快,更可扩展的方法需要最小的细胞,并改进了现有的PRO-seq协议.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 文字转录学 (Transcriptomics) 是一个学科.
背景情况:
- 精密核运行 (PRO) 测序 (PRO-seq) 地图聚合酶活动和新生的转录.
- 目前的PRO-seq协议是漫长的,复杂的,需要大量的起始材料.
研究的目的:
- 开发一个更快,更高效的 PRO-seq 协议 (rPRO-seq).
- 克服现有的PRO-seq方法在时间,技术难度和样本输入方面的局限性.
主要方法:
- 开发了快速的PRO-seq (rPRO-seq),使用预基化单链DNA (AppDNA) 和二聚体阻断寡核酸 (DBO).
- 包含在珠子上的5'RNA末端修复和基于列的净化.
- rPRO-seq可在大约12小时内进行转录组映射.
主要成果:
- rPRO-seq显著减少了协议时间,并提高了绑定效率.
- 取消了适配器二度体,并将样品损失/RNA降解降至最低.
- rPRO-seq的可复制性与标准的PRO-seq相当,可扩展到25,000个细胞.
结论:
- rPRO-seq是一种强大的,高效的,可扩展的新生转录分析方法.
- 应用了rPRO-seq以确定 *Ints11* 在小鼠造血干细胞和原始细胞恒温中的作用.
- rPRO-seq 便于以最小的样本输入进行患者特定的全基因组转录概况.
相关概念视频
03:21RNA-seq
9.9K
RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
9.9K
02:24Ribosome Profiling
3.5K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
3.5K