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基于组合DNA条形码微珠的微剖面组织样本的多重转录组NGS库的构建

Kaitong Dang¹, Yue Zhao¹, Kaiqiang Ye¹

¹State Key Laboratory of Bioelectronics, School of Biological Science & Medical Engineering, Southeast University, Nanjing, China.

Biotechnology journal

|October 11, 2023

概括

我们开发了组合DNA条形码 (CDB) -seq,这是一个具有成本效益的空间转录学方法. 这种技术可以对微切割组织进行中等通量转录组分析,解决当前高通量方法的局限性.

科学领域:

基因组学就是基因组学.
分子生物学分子生物学
生物技术是生物技术.

背景情况:

空间转录组学可以使用单细胞RNA测序和微剖析进行定位转录组分析.
由于高成本和时间要求,目前的方法面临着吞吐量方面的挑战,特别是在单细胞水平.

研究的目的:

开发一种中等通量,多重复合的方法,用于微切割组织样本的空间转录组分析.
为了解决现有的高通量空间转录学技术的局限性.

主要方法:

拟议的组合DNA条形码 (CDB) -seq,集成Smart-3SEQ和CDB磁性微珠.
优化了CDB微珠制备条件,并将该方法应用于各种RNA提取物和组织类型 (新鲜冷和FFPE小鼠大脑).

主要成果:

通过使用CDB-seq.成功地从几十个微切割样本中获得了转录组形状.
展示了一个简单的,具有成本效益的空间转录组分析工作流程.

结论:

CDB-seq为空间转录组学提供了一个有希望的,易于使用的方法.
该技术可促进微切割组织的中等吞吐量分析,扩大研究能力.

关键词:

组合DNA条形码是一个组合DNA条形码.甲固态氨酸嵌入的样品新鲜的冷组织.多重复杂的多重复杂.转录组 (transcriptome) 是一个转录组.

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RNA-seq

RNA-seq

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...