Related Experiment Video
Updated: Jan 14, 2026
09:40
Identification of Coding and Non-coding RNA Classes Expressed in Swine Whole Blood
Published on: November 28, 2018
7.7K
Technical Considerations for Blood RNA Sequencing in Genetic Testing: Evaluation of Globin Depletion Methods, Batch
Xinyi Lu1, Yumeng Ma1, Xiaomei Luo1
1Department of Clinical Genetics Center, Shanghai Institute for Pediatric Research, Xinhua Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Annals of Laboratory Medicine
|January 12, 2026
Summary
Reagent-based globin removal is effective for clinical whole-blood RNA sequencing, offering better gene detection and requiring fewer reads. Peripheral blood mononuclear cells (PBMCs) are the preferred sample type for genetic diagnostics.
Area of Science:
- Genomics
- Molecular Biology
- Clinical Diagnostics
Background:
- Blood RNA sequencing (RNA-seq) is crucial for diagnosing genetic disorders, but requires technical optimization for clinical use.
- Key technical factors for blood RNA-seq include globin depletion, batch variability, and sample type (whole blood vs. peripheral blood mononuclear cells [PBMCs]).
Purpose of the Study:
- To evaluate reagent-based globin removal (globin-RR) versus bioinformatic globin removal (globin-BR) for whole-blood RNA-seq.
- To assess batch effects in globin-RR samples and compare whole blood versus PBMCs for RNA-seq.
- To determine the optimal methods for clinical implementation of blood RNA-seq.
Main Methods:
- Compared globin-RR and globin-BR for efficiency and sequencing depth.
- Analyzed batch effects using principal component and correlation analyses.
- Compared whole blood and PBMCs for expression correlation and detection of Mendelian disease genes.
Main Results:
- Globin-RR demonstrated superior globin depletion, better detection of Mendelian disease genes (51.85% vs. 40.79%), and required fewer reads (~107 million vs. ~149 million) compared to globin-BR.
- Globin-RR showed low batch effects (inter-batch correlation, R²>0.96).
- PBMCs yielded higher detection of Mendelian disease genes (3,675 vs. 3,598) and highly expressed genes (54.46% vs. 51.85%) compared to whole blood, making them the preferred sample type.
Conclusions:
- Globin-RR is an effective, reproducible, and practical method for clinical whole-blood RNA-seq.
- PBMCs offer high concordance and a broader gene detection range, serving as a viable alternative for genetic diagnostic tests.
- Optimized technical considerations enhance the clinical utility of blood RNA-seq for genetic disorder diagnosis.
Related Concept Videos
03:21RNA-seq
11.7K
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...
11.7K
02:24Ribosome Profiling
4.1K
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...
4.1K