Journal
/
Genetics
/
Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish

Screening Sperm for the Rapid Isolation of Germline Edits in Zebrafish

DOI:
10.3791/64686-v

•

05:55 min

•

February 10, 2023

•

, ,
1Department of Molecular Biosciences,The University of Texas at Austin, 2Department of Nutritional Sciences,The University of Texas at Austin

Chapters

  • 00:04Introduction
  • 00:50Fish Sperm Collection
  • 02:11DNA Extraction and Amplification
  • 03:14PCR Amplicon Separation by Gel Electrophoresis
  • 04:20Results: Mutation Analysis of Sperm
  • 05:16Conclusion

Summary

Automatic Translation

Automatic Translation

CRISPR-Cas technologies have revolutionized the field of genome editing. However, finding and isolating the desired germline edit remains a major bottleneck. Therefore, this protocol describes a robust method for quickly screening F0 CRISPR-injected zebrafish sperm for germline edits using standard PCR, restriction digest, and gel electrophoresis techniques.

Tags

Keywords: Sperm ScreeningGermline EditsZebrafishCRISPRF0IndelsGenome EditsIn Vitro FertilizationPCRGel ElectrophoresisAnesthesiaCloacaCapillary ActionSodium HydroxideThermal CyclerTris-HClPCR ReactionAgarose Gel

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/64686?article_type=v

Novel RNA-Binding Proteins Isolation by the RaPID Methodology
check_url/64686?article_type=v

High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii
check_url/64686?article_type=v

Detection of Copy Number Alterations Using Single Cell Sequencing
check_url/64686?article_type=v

Robust DNA Isolation and High-throughput Sequencing Library Construction for Herbarium Specimens
check_url/64686?article_type=v

Formaldehyde-assisted Isolation of Regulatory Elements to Measure Chromatin Accessibility in Mammalian Cells
check_url/64686?article_type=v

Mating-based Overexpression Library Screening in Yeast
check_url/64686?article_type=v

Isolation, Characterization and MicroRNA-based Genetic Modification of Human Dental Follicle Stem Cells
check_url/64686?article_type=v

Screening Cotton Genotypes for Reniform Nematode Resistance
check_url/64686?article_type=v

Characterizing Histone Post-translational Modification Alterations in Yeast Neurodegenerative Proteinopathy Models
check_url/64686?article_type=v

Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells

Read Article