Related Experiment Video

Updated: Jun 6, 2026

Measurement of Poly A Tail Length from Drosophila Larva Brain and Cell Line
08:16

Measurement of Poly A Tail Length from Drosophila Larva Brain and Cell Line

Published on: January 12, 2024

Fractionation of mRNA based on the length of the poly(A) tail

Hedda A Meijer1, Cornelia H de Moor

  • 1Toxicology Unit, Medical Research Council, Hodgkin Building, University of Leicester, Lancaster Road, Leicester, LE, 9HN, UK.

Methods in Molecular Biology (Clifton, N.J.)
|December 3, 2010
PubMed

Insights

Poly(A) tail fractionation separates messenger RNA (mRNA) by poly(A) tail length. This technique accurately quantifies mRNA fractions for analyzing tail length changes or preparing samples for microarray analysis.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Polyadenylation is crucial for mRNA stability and translation.
  • Poly(A) tail length variations influence gene expression.
  • Accurate methods are needed to analyze poly(A) tail length.

Purpose of the Study:

  • To present a method for poly(A) tail fractionation.
  • To enable detection of subtle changes in poly(A) tail length.
  • To facilitate sample preparation for downstream analyses like microarrays.

Main Methods:

  • RNA or lysate is incubated with biotinylated oligo(dT) and streptavidin-coated paramagnetic beads.
  • mRNA is eluted sequentially using buffers of varying salt concentrations.
  • Oligoadenylated mRNA is eluted first, followed by polyadenylated mRNA.

Main Results:

  • The method allows for precise quantification of RNA in eluted fractions.
  • It separates mRNA based on poly(A) tail length.
  • Elution in multiple fractions enables detailed poly(A) tail length analysis.

Conclusions:

  • Poly(A) fractionation is a technically simple, rapid, and reproducible technique.
  • It is highly effective for analyzing poly(A) tail length variations.
  • The method is versatile and applicable to various RNA samples.

More Related Videos

Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale10:56

Polysome Fractionation and Analysis of Mammalian Translatomes on a Genome-wide Scale

Published on: May 17, 2014

Polysome Profiling without Gradient Makers or Fractionation Systems05:56

Polysome Profiling without Gradient Makers or Fractionation Systems

Published on: June 1, 2021

Related Concept Videos

Pre-mRNA Processing: Modification of pre-mRNA Ends01:35

Pre-mRNA Processing: Modification of pre-mRNA Ends

In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps the cell...
pre-mRNA Processing02:01

pre-mRNA Processing

In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
Pre-mRNA Processing02:01

Pre-mRNA Processing

In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl guanosine). This 5’ cap helps the...
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
mRNA Stability and Gene Expression02:51

mRNA Stability and Gene Expression

The structure and stability of mRNA molecules regulates gene expression, as mRNAs are a key step in the pathway from gene to protein. In eukaryotes, the half-life of mRNA varies from a few minutes up to several days. mRNA stability is essential in growth and development. The absence of the proteins regulating its stability, such as tristetraprolin in mice, can cause systemic issues, including bone marrow overgrowth, inflammation, and autoimmunity.
Cis-acting Elements involved in mRNA stability
Regulation of Expression Occurs at Multiple Steps02:24

Regulation of Expression Occurs at Multiple Steps

Gene expression can be regulated at almost every step from gene to protein. Transcription is the step that is most commonly regulated. This involves the binding of proteins to short regulatory sequences on the DNA. This association can either promote or inhibit the transcription of a gene associated with the respective sequence.
Transcription results in the generation of precursor (pre-mRNA) that consists of both exons and introns, which needs further processing before being translated to a...