ArticleScientific reports2015
An optimised direct lysis method for gene expression studies on low cell numbers.
Article in Scientific reports, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- Development of a Direct Cell-to-PCR Lysis Buffer Using Optimized Non-Ionic Detergents for RNA-Extraction-Free RT-qPCR.Methods and protocols · 2026Article
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- Cost and time-efficient construction of a 3'-end mRNA library from unpurified bulk RNA in a single tube.Experimental & molecular medicine · 2024Article
- Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review.Biosensors · 2023Review
- High-quality single-cell transcriptomics from ovarian histological sections during folliculogenesis.Life science alliance · 2023Article
- A dual paper-based nucleic acid extraction method from blood in under ten minutes for point-of-care diagnostics.The Analyst · 2023Article
- DeLTa-Seq: direct-lysate targeted RNA-Seq from crude tissue lysate.Plant methods · 2022Article
- An Optimized Direct Lysis Gene Expression Microplate Assay and Applications for Disease, Differentiation, and Pharmacological Cell-Based Studies.Biosensors · 2022Article
- Sensitive and accurate analysis of gene expression signatures enabled by oligonucleotide-labelled cDNA.RNA biology · 2022Article
- A Simple, Cost-Effective, and Automation-Friendly Direct PCR Approach for Bacterial Community Analysis.mSystems · 2021Article
- Performance of a scalable RNA extraction-free transcriptome profiling method for adherent cultured human cells.Scientific reports · 2021Article
- Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630.Scientific reports · 2021Article
- Facile Fabrication of Thin-Bottom Round-Well Plates Using the Deformation of PDMS Molds and Their Application for Single-Cell PCR.Micromachines · 2020Article
- One step DNA amplification of mammalian cells in picoliter microwell arrays.RSC advances · 2019Article
- Microscale Gene Expression Analysis of Tumor-Associated Macrophages.Scientific reports · 2018Article
- A novel ex vivo Huntington's disease model for studying GABAergic neurons and cell grafts by laser microdissection.PloS one · 2018Article
- The cytotoxic effects of dimethyl sulfoxide in mouse preimplantation embryos: a mechanistic study.Theranostics · 2017Article
- Gene expression profiling and DNA methylation analyses of CTCs.Molecular oncology · 2016Review
- Sequential fractionation and isolation of subcellular proteins from tissue or cultured cells.MethodsX · 2015Article
Corrections and comments
- Erratum issued
Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
There is increasing interest in gene expression analysis of either single cells or limited numbers of cells. One such application is the analysis of harvested circulating tumour cells (CTCs), which are often present in very low numbers. A highly efficient protocol for RNA extraction, which involves a minimal number of steps to avoid RNA loss, is essential for low input cell numbers. We compared several lysis solutions that enable reverse transcription (RT) to be performed directly on the cell lysate, offering a simple rapid approach to minimise RNA loss for RT. The lysis solutions were assessed by reverse transcription quantitative polymerase chain reaction (RT-qPCR) in low cell numbers isolated from four breast cancer cell lines. We found that a lysis solution containing both the non-ionic detergent (IGEPAL CA-630, chemically equivalent to Nonidet P-40 or NP-40) and bovine serum albumin (BSA) gave the best RT-qPCR yield. This direct lysis to reverse transcription protocol outperformed a column-based extraction method using a commercial kit. This study demonstrates a simple, reliable, time- and cost-effective method that can be widely used in any situation where RNA needs to be prepared from low to very low cell numbers.
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