1. Female adult Xenopus laevis frogs (Xenopus-Express, Homosassa, FL, USA, see Website: http://www.xenopus.com; or NASCO Science Supply, Fort Atkinson, WI, USA, see Website: http://www.enasco.com). Frogs must be kept in dechlorinated fresh water (one frog per gallon) at 18°C with a regulated light cycle (12 h light, 12 h dark). Animals should be fed twice weekly and have their tank water changed following meals (see Note 1).
2. Anesthetic stock solution: 3% benzocaine (ethyl-p-aminobenzoate) in ethanol. Dilute 1:100 with water for use with frogs.
3. Surgical instruments including small scissors, forceps, and needle holder.
4. Collagenase (type 1A from Clostridium histolyticum, Sigma Chemical Co., cat. no. C-9891) dissolved in Ca2+ free ND-96 to final concentration of 2.5 mg/mL.
5. Ca2+ free ND-96: 96 mM NaCl, 2 mM KCl, 1 mM MgCl2, 5 mM HEPES, adjust pH to 7.5 with NaOH.
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