ArticleScientific reports2017
Global preamplification simplifies targeted mRNA quantification.
Article in Scientific reports, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
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- DigiMir Test: Establishing a Novel Pipeline for MiR-371a Quantification Using Droplet Digital PCR in Liquid Biopsies From Testicular Germ Cell Tumor Patients.Frontiers in oncology · 2022Article
- Tutorial: Guidelines for Single-Cell RT-qPCR.Cells · 2021Review
- Regulation of gene expression in the bovine blastocyst by colony-stimulating factor 2 is disrupted by CRISPR/Cas9-mediated deletion of CSF2RA.Biology of reproduction · 2021Article
- Effects of GLP-1 receptor agonist on changes in the gut bacterium and the underlying mechanisms.Scientific reports · 2021Article
- An improved method for specific-target preamplification PCR analysis of single blastocysts useful for embryo sexing and high-throughput gene expression analysis.Journal of dairy science · 2021Article
- Non-Invasive Diagnosis for Acute Rejection Using Urinary mRNA Signature Reflecting Allograft Status in Kidney Transplantation.Frontiers in immunology · 2021Article
- Development and validation of a 25-Gene Panel urine test for prostate cancer diagnosis and potential treatment follow-up.BMC medicine · 2020Article
- Article
- Targeted transcript quantification in single disseminated cancer cells after whole transcriptome amplification.PloS one · 2019Article
- Preamplification with dUTP and Cod UNG Enables Elimination of Contaminating Amplicons.International journal of molecular sciences · 2018Article
- Unravelling the biological secrets of microchimerism by single-cell analysis.Briefings in functional genomics · 2018Article
- Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization.Journal of visualized experiments : JoVE · 2018Article
- Article
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Authors and funding
5 authors.
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Abstract
The need to perform gene expression profiling using next generation sequencing and quantitative real-time PCR (qPCR) on small sample sizes and single cells is rapidly expanding. However, to analyse few molecules, preamplification is required. Here, we studied global and target-specific preamplification using 96 optimised qPCR assays. To evaluate the preamplification strategies, we monitored the reactions in real-time using SYBR Green I detection chemistry followed by melting curve analysis. Next, we compared yield and reproducibility of global preamplification to that of target-specific preamplification by qPCR using the same amount of total RNA. Global preamplification generated 9.3-fold lower yield and 1.6-fold lower reproducibility than target-specific preamplification. However, the performance of global preamplification is sufficient for most downstream applications and offers several advantages over target-specific preamplification. To demonstrate the potential of global preamplification we analysed the expression of 15 genes in 60 single cells. In conclusion, we show that global preamplification simplifies targeted gene expression profiling of small sample sizes by a flexible workflow. We outline the pros and cons for global preamplification compared to target-specific preamplification.
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