Transgenesis in Xenopus laevis
Improvements to the transgenic technique

We have been using the transgenic procedure developed by Kroll and Amaya to produce transgenic Xenopus laevis. We encountered many problems with the procedure over the course of our experiments. Many of the problems (and solutions) are addressed by Kroll and Amaya, both in their protocol and in the Troubleshooting and Q&A sections of their web page (http://www.welc.cam.ac.uk/~ea3/Trouble.Shooting.Section.html and http://www.welc.cam.ac.uk/~ea3/Ques.Ans.Section.htm). We have found that additional modifications to the technique improves the efficiency of transgenesis. Here are some more useful improvements that we discovered the hard way.

Transgenic reaction mixture

               - Use cut pipet tips and pipet slowly.
               - Don't get bubbles in the solution when pipetting
               - Keep condensed nuclei on ice, but keep decondensed nuclei at room temperature.

Transplantation needles

Agarose dishes Sperm preparation Egg collection and dejellying Microinjecting Monitoring embryos  This page created by members of the Browder Lab: Rob Chan, Ivana Kostic and Pierrette Lo.
 July 29, 1999