Features Partner Sites Information LinkXpress hp
Sign In
Advertise with Us
LGC Clinical Diagnostics

Download Mobile App




RNA Sequencing Offers Greater Capacity for Recovery and Analysis of Molecules

By LabMedica International staff writers
Posted on 01 Jan 2019
Print article
Image: The Nadia automated single cell Drop-seq RNA sequencing system (Photo courtesy of Dolomite Bio).
Image: The Nadia automated single cell Drop-seq RNA sequencing system (Photo courtesy of Dolomite Bio).
A recent paper described a significant modification that enables Drop-seq single-cell RNA sequencing technology to recover and analyze a more diverse variety of molecules.

Drop-seq methodology involves encapsulating single cells with single barcoded beads in nanoliter-sized droplets. The barcoded oligo bead library is constructed such that each bead has a unique DNA barcode sequence, but within a bead, the thousands of copies of oligo all contain an identical barcode sequence. The 3′ end of the oligo has a poly(dT) stretch that captures messenger RNA (mRNA) and primes reverse transcription. Once encapsulated, the cell is broken open and the mRNA is captured on the bead, resulting in single-cell transcriptomes attached to microparticles. The RNA is converted to DNA, amplified and sequenced. The major drawback to the technique is that it can only identify molecules of mRNA, which limits the potential scope of analyses.

Investigators at Cornell University (Ithaca, NY, USA) described in the December 17, 2018, online edition of the journal Nature Methods a modification to Drop-seq. Their DART-seq (droplet-assisted RNA targeting by single-cell sequencing) method was depicted as being a versatile technology that enabled multiplexed amplicon sequencing and transcriptome profiling in single cells. The modification was accomplished by enzymatically customizing the beads prior to performing conventional Drop-seq analysis, which allowed for the recovery and analysis of a greater variety of molecules.

The investigators applied DART-seq to simultaneously characterize the non-A-tailed transcripts of a segmented dsRNA virus and the transcriptome of the infected cell. In addition, they used DART-seq to simultaneously determine the natively paired, variable region heavy and light chain amplicons and the transcriptome of B-lymphocytes.

"Those technologies are very popular because they have lowered the cost of these types of analyses and sort of democratized them, made them very cheap and easy to do for many labs," said senior author Dr. Iwijin De Vlaminck, assistant professor in of biomedical engineering at Cornell University.

Related Links:
Cornell University

Gold Member
Troponin T QC
Troponin T Quality Control
Verification Panels for Assay Development & QC
Seroconversion Panels
New
Respiratory Bacterial Panel
Real Respiratory Bacterial Panel 2
New
TORCH Infections Test
TORCH Panel

Print article

Channels

Clinical Chemistry

view channel
Image: The tiny clay-based materials can be customized for a range of medical applications (Photo courtesy of Angira Roy and Sam O’Keefe)

‘Brilliantly Luminous’ Nanoscale Chemical Tool to Improve Disease Detection

Thousands of commercially available glowing molecules known as fluorophores are commonly used in medical imaging, disease detection, biomarker tagging, and chemical analysis. They are also integral in... Read more

Immunology

view channel
Image: The cancer stem cell test can accurately choose more effective treatments (Photo courtesy of University of Cincinnati)

Stem Cell Test Predicts Treatment Outcome for Patients with Platinum-Resistant Ovarian Cancer

Epithelial ovarian cancer frequently responds to chemotherapy initially, but eventually, the tumor develops resistance to the therapy, leading to regrowth. This resistance is partially due to the activation... Read more

Microbiology

view channel
Image: The lab-in-tube assay could improve TB diagnoses in rural or resource-limited areas (Photo courtesy of Kenny Lass/Tulane University)

Handheld Device Delivers Low-Cost TB Results in Less Than One Hour

Tuberculosis (TB) remains the deadliest infectious disease globally, affecting an estimated 10 million people annually. In 2021, about 4.2 million TB cases went undiagnosed or unreported, mainly due to... Read more

Pathology

view channel
Image: The ready-to-use DUB enzyme assay kits accelerate routine DUB activity assays without compromising data quality (Photo courtesy of Adobe Stock)

Sensitive and Specific DUB Enzyme Assay Kits Require Minimal Setup Without Substrate Preparation

Ubiquitination and deubiquitination are two important physiological processes in the ubiquitin-proteasome system, responsible for protein degradation in cells. Deubiquitinating (DUB) enzymes contain around... Read more

Technology

view channel
Image: The HIV-1 self-testing chip will be capable of selectively detecting HIV in whole blood samples (Photo courtesy of Shutterstock)

Disposable Microchip Technology Could Selectively Detect HIV in Whole Blood Samples

As of the end of 2023, approximately 40 million people globally were living with HIV, and around 630,000 individuals died from AIDS-related illnesses that same year. Despite a substantial decline in deaths... Read more

Industry

view channel
Image: The collaboration aims to leverage Oxford Nanopore\'s sequencing platform and Cepheid\'s GeneXpert system to advance the field of sequencing for infectious diseases (Photo courtesy of Cepheid)

Cepheid and Oxford Nanopore Technologies Partner on Advancing Automated Sequencing-Based Solutions

Cepheid (Sunnyvale, CA, USA), a leading molecular diagnostics company, and Oxford Nanopore Technologies (Oxford, UK), the company behind a new generation of sequencing-based molecular analysis technologies,... Read more
Copyright © 2000-2025 Globetech Media. All rights reserved.