Protocol

Tissue Sampling and Genomic DNA Purification from the Western Clawed Frog Xenopus tropicalis

¹ UNC McAllister Heart Institute, Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
² Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA
³ Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA

⁴Corresponding author (chris_showell{at}med.unc.edu).

INTRODUCTION

The extraction of genomic DNA from Western clawed frog (Xenopus tropicalis) tissue samples is an essential step for genotyping known mutations, identifying live animals carrying nonfluorescent transgenes, and reverse genetic screening techniques. We describe here a method for sampling tail tissue from X. tropicalis tadpoles at stage 48 and later. This method of tissue sampling allows a significant amount of genomic DNA to be obtained from tadpoles without killing them. In both Xenopus laevis and X. tropicalis, the tip of the tail is able to regenerate following surgery and its removal does not have a significant effect on the survival of the tadpole when performed carefully. Genomic DNA purification can be carried out in "deep-well" 96-well plates, making it amenable to high-throughput applications. Typically, the DNA yield is sufficient for more than 100 standard 50-µL polymerase chain reactions (PCRs) (using 1-5 µL of DNA per reaction), so it can be used for genetic screening and mapping studies.