Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

66.5K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
66.5K
RNA Splicing01:32

RNA Splicing

61.3K
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
61.3K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Transient receptor potential melastatin 3 ion channel expressed in sensory neurons mediates osteoarthritis pain in mice.

Osteoarthritis and cartilage·2026
Same author

Regulation of ADAM10 activity through microdomain-dependent intracellular calcium changes.

Cell communication and signaling : CCS·2024
Same author

Changes of Protein Expression after CRISPR/Cas9 Knockout of miRNA-142 in Cell Lines Derived from Diffuse Large B-Cell Lymphoma.

Cancers·2022
Same author

Control of TRPM3 Ion Channels by Protein Kinase CK2-Mediated Phosphorylation in Pancreatic β-Cells of the Line INS-1.

International journal of molecular sciences·2021
Same author

Prolactin-sensitive olfactory sensory neurons regulate male preference in female mice by modulating responses to chemosensory cues.

Science advances·2021
Same author

Cavβ3 Regulates Ca<sup>2+</sup> Signaling and Insulin Expression in Pancreatic β-Cells in a Cell-Autonomous Manner.

Diabetes·2021

Related Experiment Video

Updated: Mar 24, 2026

Merging Absolute and Relative Quantitative PCR Data to Quantify STAT3 Splice Variant Transcripts
11:19

Merging Absolute and Relative Quantitative PCR Data to Quantify STAT3 Splice Variant Transcripts

Published on: October 9, 2016

15.7K

A reliable method for quantification of splice variants using RT-qPCR.

Julia Camacho Londoño1, Stephan E Philipp2

  • 1Experimentelle und Klinische Pharmakologie und Toxikologie, Universität des Saarlandes, 66421, Homburg, Germany.

BMC Molecular Biology
|March 17, 2016
PubMed
Summary
This summary is machine-generated.

This study introduces a new, reliable method for quantifying alternative splicing variants using one-step RT-qPCR. This technique accurately measures splice variant incidence, aiding in understanding RNA splicing events.

More Related Videos

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models
09:58

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

Published on: December 9, 2016

14.5K
Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
11:48

Detection of Alternative Splicing During Epithelial-Mesenchymal Transition

Published on: October 9, 2014

13.5K

Related Experiment Videos

Last Updated: Mar 24, 2026

Merging Absolute and Relative Quantitative PCR Data to Quantify STAT3 Splice Variant Transcripts
11:19

Merging Absolute and Relative Quantitative PCR Data to Quantify STAT3 Splice Variant Transcripts

Published on: October 9, 2016

15.7K
Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models
09:58

Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models

Published on: December 9, 2016

14.5K
Detection of Alternative Splicing During Epithelial-Mesenchymal Transition
11:48

Detection of Alternative Splicing During Epithelial-Mesenchymal Transition

Published on: October 9, 2014

13.5K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Protein isoforms commonly result from alternative RNA splicing.
  • Accurate quantification of single splicing events is crucial but challenging with existing methods.
  • Current techniques for assessing splicing incidence are often laborious, unreliable, or have limited applicability.

Purpose of the Study:

  • To develop and validate an improved method for determining the relative incidence of alternative splice variants at a single site.
  • To provide a reliable and precise technique for quantifying splice variants within a single sample.

Main Methods:

  • Utilized one-step reverse transcription quantitative PCR (RT-qPCR) for splice variant quantification.
  • Employed variant-specific and common primer pairs for comparative amplification.
  • Verified amplicon identity using melt curve analysis.
  • Incorporated an internal control (sum of incidences = 100%) to monitor experimental accuracy and reverse transcription uniformity.

Main Results:

  • The developed method accurately quantifies relative splice variant incidence in a single sample.
  • Melt curve analysis confirmed the identity of variant-specific amplicons.
  • The internal control feature effectively monitored experimental errors and reverse transcription consistency.
  • Validation using cDNA mixtures and diverse RNA samples demonstrated the method's reliability.

Conclusions:

  • The novel RT-qPCR method is easy to implement and does not require reference genes or standard curves.
  • It offers a reliable and precise approach for distinguishing minor differences in the incidence of two splice variants.
  • This technique overcomes limitations of existing labor-intensive and error-prone methods.