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Genotyping Transgenic Zebrafish Using Genomic DNA Extracted from Clutch of Embryos

LJ Lili Jing

Published: Feb 5, 2012 DOI: 10.21769/BioProtoc.181 Views: 14622

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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

  1. Zebrafish embryos
  2. MeOH
  3. Phenol
  4. Chloroform
  5. Isoamyl alcohol (IAA)
  6. NaCl
  7. KCl
  8. MgCl2
  9. EtOH
  10. TE
  11. Tween 20
  12. NaOAC
  13. Gelatine
  14. NP40
  15. Proteinase K
  16. ddH2O
  17. Phenol: chloroform: isoamyl alcohol (25:24:1)
  18. Primers (custom ordered from IDT)
  19. 1x RAPD buffer (see Recipes)
  20. RAPD+ (see Recipes)
  21. PCR lysis buffer (see Recipes)

Equipment

  1. PCR thermal cycler
  2. Incubator
  3. Glass pipette
  4. 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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