Evidence map›Paper›PMID 29021971›Full record

ArticleBiomolecular detection and quantification2017

Evaluation of relative quantification of alternatively spliced transcripts using droplet digital PCR.

Mattias Van Heetvelde, Wouter Van Loocke, Wim Trypsteen, Annelot Baert, Katrien Vanderheyden, Brecht Crombez, Jo Vandesompele, Kim De Leeneer, Kathleen B M Claes

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In one paragraph

Article in Biomolecular detection and quantification, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.

0numbers the graph read from it
0cells of the map it votes in
11citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

11 citing papers in PubMed.

  1. Article
  2. Article
  3. Review
  4. Article
  5. Article
  6. Article
  7. Article
  8. Review
  9. Article
  10. BRCA1-No Matter How You Splice It.Cancer research · 2019
    Review
  11. Non-Coding Variants inCancers · 2018
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

9 authors.

Mattias Van HeetveldeCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Wouter Van LoockeCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Wim TrypsteenHIV Translational Research unit, Department of Internal Medicine, Ghent University, Ghent, Belgium.
Annelot BaertDepartment of Basic Medical Sciences, Ghent University, Ghent, Belgium.
Katrien VanderheydenCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Brecht CrombezCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Jo VandesompeleCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Kim De LeeneerCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.
Kathleen B M ClaesCenter for Medical Genetics Ghent, Ghent University Hospital, Ghent, Belgium.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

introductionFor the relative quantification of isoform expression, RT-qPCR has been the gold standard for over a decade. More recently, digital PCR is becoming widely implemented, as it is promised to be more accurate, sensitive and less affected by inhibitors, without the need for standard curves. In this study we evaluated RT-qPCR versus RT-droplet digital PCR (ddPCR) for the relative quantification of isoforms in controls and carriers of the splice site mutation MATERIALS AND

methodsRNA was extracted from EBV cell lines of controls and heterozygous

resultsBoth ddPCR and RT-qPCR were able to accurately quantify all targets and showed the same LOB, LOD and LOQ; also precision and reproducibility were similar. Both techniques have the same dynamic range and linearity at biologically relevant template concentrations. However, a significantly higher cost and workload was required for ddPCR experiments.

conclusionsOur study recognizes the potential and validity of digital PCR but shows the value of a highly optimized qPCR for the relative quantification of isoforms. Cost efficiency and simplicity turned out to be better for RT-qPCR.

Indexed as

Alternative splicingDroplet digital PCRReverse transcriptase polymerase chain reaction

Identifiers

PMID29021971
PMCPMC5634819

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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.