Rivista
/
Genetica
/
CRISPR Guide RNA Cloning for Mammalian Systems
JoVE Journal
Genetica
This content is Free Access.
JoVE Journal Genetica
CRISPR Guide RNA Cloning for Mammalian Systems

CRISPR Guide RNA Cloning for Mammalian Systems

DOI:
10.3791/57998-v

•

06:48 min

•

October 02, 2018

•

, , , , , , ,
1Wyss Institute for Biologically Inspired Engineering,Harvard University, 2Department of Genetics,Harvard Medical School, 3Department of Pathology,Massachusetts General Hospital, 4Institute for Medical Engineering & Science,Massachusetts Institute of Technology, 5Synthetic Biology Center,Massachusetts Institute of Technology, 6Department of Biological Engineering,Massachusetts Institute of Technology, 7Broad Institute

Capitoli

  • 00:04Titolo
  • 00:25Preparation of the Primers and Expression Vectors
  • 02:05Ligating the Annealed gRNA Oligonucleotides
  • 02:59Transformation of E. coli
  • 04:03PCR Analysis
  • 05:35Results: CRISPR Guided RNA Cloning
  • 06:23Conclusion

Summary

Traduzione automatica

Traduzione automatica

Here, a simple, efficient, and cost-effective method of sgRNA cloning is outlined.

Tags

CRISPRGuide RNACloningMammalian SystemPlasmidSingle-guide RNAOligonucleotideLigationE. ColiCompetent CellsTransformation

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

Mapping RNA-RNA Interactions Globally Using Biotinylated Psoralen

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

CRISPR-Mediated Reorganization of Chromatin Loop Structure

Rare Event Detection Using Error-corrected DNA and RNA Sequencing

Endogenous Protein Tagging in Human Induced Pluripotent Stem Cells Using CRISPR/Cas9

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Artificial RNA Polymerase II Elongation Complexes for Dissecting Co-transcriptional RNA Processing Events

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

Read Article