概要: Frontal Cortex whole transcriptome sequencing in COVID-19 elderly patients
项目整体设计: SARS-Cov-2 infection is frequently associated with Nervous System (NS) manifestations. However, it is not clear how SARS-CoV-2 can cause neurological dysfunctions and which molecular processes affects. In this work, we examined the frontal cortex tissue of patients who died of COVID-19 for the presence of SARS-CoV-2, comparing qRT-PCR with ddPCR. We also investigated the transcriptomic profile of frontal cortex from COVID-19 patients and matched controls by RNA-seq analysis to characterize the transcriptional signature.
Autoptic human brain samples were collected. Slices of anterior frontal lobe were fresh cut into about 10 mm thickness and frozen at -80°C.
处理方案:
-
提取方案:
Samples were homogenized and total RNA was isolated by Trizol® reagent (Life Science Technologies, Italy) following the manufacturer’s specifications. RNAs were quantified using a Nanodrop ND-100 Spectrophotometer (Nanodrop Technologies, Wilmington, USA) and a 2100 Bioanalyzer (Agilent RNA 6000 Nano Kit, Waldbronn, Germany); RNAs with a 260:280 ratio of ≥1.5 and an RNA integrity number of ≥8 were subjected to deep sequencing.
Sequencing libraries were prepared by the Lexogen CORALL Total RNA-Seq Library Prep Kit using 500-ng total RNA (Lexogen). Qualities of sequencing libraries were assessed with 4200 TapeStation with the DNA1000 reagent kit. RNA processing has been carried out using Illumina NextSeq 500 Sequencing.
建库方案:
Library Prep Kit using 500-ng total RNA (Lexogen). Qualities of sequencing libraries were assessed with 4200 TapeStation with the DNA1000 reagent kit. RNA processing has been carried out using Illumina NextSeq 500 Sequencing.
测序信息
分子类型:
total RNA
库的片段类型:
PAIRED
库的链类型:
Reverse; -
测序平台:
ILLUMINA
测序仪型号:
Illumina NextSeq 500
链特异性:
Specific; Unspecific
样本
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数据来源
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组织
细胞类型
细胞亚型
细胞系
疾病
疾病状态
发育阶段
突变/变异
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Condition Detail
生长方案
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分子类型
库的片段类型
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加标(Spike-In)
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平均测序片段长度_1
平均测序片段长度_2
唯一比对率
多重比对率
覆盖度
文章
Multi-omic profiling reveals widespread dysregulation of innate immunity and hematopoiesis in COVID-19.
The Journal of experimental medicine . 2021-06-15 [PMID:
34128959]
Detection of SARS-CoV-2 genome and whole transcriptome sequencing in frontal cortex of COVID-19 patients.
Brain, behavior, and immunity . 2021-05-19 [PMID:
34022369]