Genotyping by skin swabbing
Skin swabbing
(zebrafish adult)
1. Add 400 µl of DNA collection buffer (30 mM Tris-HCl
and 1 μg/μL PK
[03115828001; Roche]) per tube.
2. Transfer zebrafish to a mating cage containing E3
embryo medium (5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2, and 0.33 mM MgSO4)
3. Anaesthetise zebrafish in 1X Tricaine E3 embryo
medium. Transfer one fish at a time onto an upsidedown petri dish using a
spoon. Remove excess water blot on paper towels, then swab several times on the
caudal fin with a cotton swab (Q-tips® Cotton Swabs) until the tip of the swab
became yellowish.
4. Cut the cotton swab 1 cm above the cotton wad, and put
into the tube in step 1 and the fish into a single box.
5. Vortex for 10 s and incubated at 98°C for 5 min
6. Remove the cotton wad, then use 5.9 µl to set up PCR reaction in a
total volume of 10 µl.
Reference: Jang JY, Liang T, Kim MK, Kang KW, Lee B, Choi SY. A Rapid and Noninvasive Method That Extracts Polymerase Chain Reaction-Ready Genomic DNA from Adult Zebrafish. Zebrafish. 2022 Aug;19(4):160-164.