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Related Concept Videos

RNA Splicing01:32

RNA Splicing

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...
DNA Agarose Gel Electrophoresis02:35

DNA Agarose Gel Electrophoresis

Agarose gel electrophoresis is a laboratory technique commonly used to separate DNA fragments by size. However, it can also be used to isolate and purify DNA fragments using a gel extraction protocol.
Gel extraction follows five major steps: running gel electrophoresis to separate fragments, isolating the individual bands, extracting DNA from those bands, and removing the dye and salts from the extracted mixture to obtain pure DNA.
In cloning experiments, both the insert and vector DNA...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Pre-mRNA Processing: RNA Splicing01:32

Pre-mRNA Processing: RNA Splicing

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...

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Related Experiment Video

Updated: May 7, 2026

Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes
05:44

Utilization of Grafix for the Detection of Transient Interactors of Saccharomyces cerevisiae Spliceosome Subcomplexes

Published on: November 9, 2020

Analysis of splicing complexes on native gels.

Manuel Ares

    Cold Spring Harbor Protocols
    |October 3, 2013
    PubMed
    Summary
    This summary is machine-generated.

    This study presents a gel-based method to analyze yeast splicing complexes. It visualizes RNA-protein interactions during pre-mRNA splicing, offering insights into this fundamental biological process.

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    Last Updated: May 7, 2026

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    A Reporter Based Cellular Assay for Monitoring Splicing Efficiency
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    Published on: September 15, 2021

    Area of Science:

    • Molecular Biology
    • Biochemistry
    • Genetics

    Background:

    • RNA splicing is a complex, ATP-dependent process involving macromolecular associations.
    • Understanding splicing requires analyzing the entire complex, not just the chemical reactions.
    • Native gel electrophoresis is a valuable technique for studying RNA-protein interactions.

    Purpose of the Study:

    • To describe a gel-based protocol for detecting and analyzing yeast splicing complexes.
    • To investigate the composition of splicing complexes formed in vitro.
    • To provide a method for studying the dynamic assembly of splicing machinery.

    Main Methods:

    • Radiolabeling of pre-mRNA substrate for detection.
    • In vitro assembly of splicing complexes.
    • Quenching of splicing reactions by dilution and competitor RNA.
    • Analysis using composite agarose-acrylamide gel electrophoresis in glycerol.
    • Detection and extraction of labeled RNA-protein complexes.

    Main Results:

    • The protocol successfully detects and analyzes yeast splicing complexes.
    • Extracted complexes contain small nuclear RNAs (snRNAs) and partially spliced pre-mRNA.
    • The method allows visualization of complexes before complete reaction.

    Conclusions:

    • This gel method provides a robust approach to study RNA splicing complexes in yeast.
    • The findings highlight the importance of analyzing the entire splicing machinery.
    • The protocol facilitates research into the dynamics of pre-mRNA splicing.