Journal
/
Biochemistry
/
MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria

DOI:
10.3791/61731-v

•

08:34 min

•

February 23, 2021

•

, , , , ,
1Université de Strasbourg, CNRS, 2Department of Biochemistry,Université de Sherbrooke

Chapters

  • 00:05Introduction
  • 01:05Bacteria Harvesting and Lysis
  • 02:52MS2-affinity Purification
  • 03:55RNA Extraction of Collected Fractions (CE, FT, W and E)
  • 05:24Results: Constructs Validation and MAPS Controls
  • 07:49Conclusion

Summary

Automatic Translation

Automatic Translation

MAPS technology has been developed to scrutinize the targetome of a specific regulatory RNA in vivo. The sRNA of interest is tagged with a MS2 aptamer enabling the co-purification of its RNA partners and their identification by RNA sequencing. This modified protocol is particularly suited for Gram-positive bacteria. 

Tags

Keywords: MS2-affinity PurificationRNA SequencingMAPSGram-positive BacteriaSRNARegulatory NetworksVirulence FactorsBiofilm FormationStaphylococcal InfectionsCell LysisAmylose ResinMBP-MS2 ProteinFlow-through Fraction

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/61731?article_type=v

Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to S-citalopram
check_url/61731?article_type=v

Growing Protein Crystals with Distinct Dimensions Using Automated Crystallization Coupled with In Situ Dynamic Light Scattering
check_url/61731?article_type=v

Single-step Purification of Macromolecular Complexes Using RNA Attached to Biotin and a Photo-cleavable Linker
check_url/61731?article_type=v

Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli
check_url/61731?article_type=v

Translating Ribosome Affinity Purification (TRAP) for RNA Isolation from Endothelial Cells In Vivo
check_url/61731?article_type=v

Extremely Rapid and Specific Metabolic Labelling of RNA In Vivo with 4-Thiouracil (Ers4tU)
check_url/61731?article_type=v

Quantitative Analysis of the Cellular Lipidome of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
check_url/61731?article_type=v

Rapid and Cost-Effective RNA Extraction of Rat Pancreatic Tissue
check_url/61731?article_type=v

Quantitative Metabolomics of Saccharomyces Cerevisiae Using Liquid Chromatography Coupled with Tandem Mass Spectrometry
check_url/61731?article_type=v

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing

Read Article