Since the tyrosinase gene is involved in melanin pigment synthesis, successful knockout of this gene should cause an absence of black pigment and consequently, albinism. Generally, the CRISPR/Cas9 method, when used on Xenopus tropicalis, does not disrupt the targeted gene in all cells, resulting in a chimeric individual with both wild-type and knockout cells. This kit contains a fixative, a color-developing solution, and X-gal as a chromogenic substrate. These kit components are necessary for visualizing the expression of beta-galactosidase. Also included is a gRNA for targeting the tyrosinase gene, but Cas9 is not included. The kit includes all the solutions described below. You may download the instruction manual here.

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  1. 1.Mixture of gRNA for tyrosinase and mRNA for nuclear-localized β-galactosidase, 2.5 µl x 2 (100 ng/µl of each RNA)

  2. 2.Fixative solution containing 2% formaldehyde, 2 mM MgCl2, 1.25 mM EGTA, 0.1 mM HEPES (pH 7.4), and 2.5 mM NaOH, 1.2 ml x 2 tubes

  3. 3.Color-developing solution containing 0.7x PBS, 2 mM MgCl2, 0.02% NP-40, 0.01% sodium deoxycholate, 5 mM K3[Fe(CN)6], and 5 mM K4[Fe(CN)6]), 1.2 ml x 2 tubes

  4. 4.2% X-gal in dimethylformamid, 120 µl