Welcome to the transgenics web site of the Cepko, Tabin, Dymecki, and Dietrich Labs. For smooth addition to the queue and most probable injection success, be sure to print and fill out your application completely and hold fast to the prescribed DNA purification protocol. Upon approval of your application you will be notified and can then provide your DNA for injection. Documentation of succesful vector separation and concentration is required. Provide a photograph of a comparative gel that includes a ladder, your fragment and other DNA bands of known concentration. See Figure 1 below.

A proven screening strategy for detection of a single transgene copy is also required. Assay by either PCR, southern blot, or slot blot. Amplify or hybridize your fragment in the context of wt tail DNA and compare to a negative control tail DNA. One copy of a 5-10kb sequence per diploid mouse genome (6x10⁹ bp) is roughly one part in one million. A standard curve should be generated with increments of 1pg, 10pg, and 100pg of transgene per 1 ug of wt tail DNA. Please be sure to provide a photo of your results. See Figure 2 below.

Completion of these requirements will result in your project’s addition to the queue. Your gene construct will be assigned two injection dates unless other parameters are negotiated with Bill Dietrich. Actual injections will take place within a week of the assigned dates. Expect pups 20 days from the injection date and tail samples within 25 days of birth. Timely screening will allow for faster allocation of additional injection days if needed.