PUBLICATION
Total Nucleic Acid Extraction from Single Zebrafish Embryos for Genotyping and RNA-seq
- Authors
- Wali, N., Merteroglu, M., White, R.J., Busch-Nentwich, E.M.
- ID
- ZDB-PUB-220205-7
- Date
- 2022
- Source
- Bio-protocol 12: e4284 (Journal)
- Registered Authors
- Busch-Nentwich, Elisabeth
- Keywords
- Genotyping, RNA-seq, Transcriptomics, Zebrafish
- MeSH Terms
- none
- PubMed
- 35118175 Full text @ Bio Protoc
Citation
Wali, N., Merteroglu, M., White, R.J., Busch-Nentwich, E.M. (2022) Total Nucleic Acid Extraction from Single Zebrafish Embryos for Genotyping and RNA-seq. Bio-protocol. 12:e4284.
Abstract
RNA sequencing allows for the quantification of the transcriptome of embryos to investigate transcriptional responses to various perturbations (e.g., mutations, infections, drug treatments). Previous protocols either lack the option to genotype individual samples, or are laborious and therefore difficult to do at a large scale. We have developed a protocol to extract total nucleic acid from individual zebrafish embryos. Individual embryos are lysed in 96-well plates and nucleic acid is extracted using SPRI beads. The total nucleic acid can be genotyped and then DNase I treated to produce RNA samples for sequencing. This protocol allows for processing large numbers of individual samples, with the ability to genotype each sample, which makes it possible to undertake transcriptomic analysis on mutants at timepoints before the phenotype is visible. Graphic abstract: Extraction of total nucleic acid from individual zebrafish embryos for genotyping and RNA-seq.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping