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Genotyping Transgenic Zebrafish Using Genomic DNA Extracted from Clutch of Embryos
Published: Feb 5, 2012 DOI: 10.21769/BioProtoc.181 Views: 14713
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Abstract
Transgenic zebrafish are very useful genetic tools to study various biological processes. Identification the right transgene founder and the subsequent transgenic animals are always tedious and time consuming. This protocol provides a relatively rapid and easy method to identify the founder parent using a clutch of embryos.
Materials and Reagents
- Zebrafish embryos
- MeOH
- Phenol
- Chloroform
- Isoamyl alcohol (IAA)
- NaCl
- KCl
- MgCl2
- EtOH
- TE
- Tween 20
- NaOAC
- Gelatine
- NP40
- Proteinase K
- ddH2O
- Phenol: chloroform: isoamyl alcohol (25:24:1)
- Primers (custom ordered from IDT)
- 1x RAPD buffer (see Recipes)
- RAPD+ (see Recipes)
- PCR lysis buffer (see Recipes)
Equipment
- PCR thermal cycler
- Incubator
- Glass pipette
- Remove MeOH and dry embryos in 55 °C incubator.
Procedure
Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.
Category
Molecular Biology > DNA > Genotyping
Molecular Biology > DNA > DNA extraction
Molecular Biology > DNA > PCR
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