Experimental flow:

1. The PI submits the appropriate service form to the TMESCSR, including all required documentation, as described on the submission form.
2. The BAC DNA purification is scheduled by the TMESCSR.  Dr. Mortlock notifies the PI of the start date.
3. The investigator provides a freshly streaked plate with isolated colonies of the BAC clone to be prepped.
4. The TMESCSR will perform a Nucleobond BAC 100 column-based DNA prep.  A sample of the DNA prep will be run on a pulsed-field gel to verify purification of intact high-molecular weight BAC DNA.  Dr. Mortlock will provide the P.I. with the purified BAC DNA dissolved in 100 µl of 10 mM Tris / 0.1 mM EDTA (“Low TE”) and a copy of the gel image.   Typical yields are 10-100 µg.  However, for some BAC clones it can be difficult to achieve these yields of DNA, so minimum yields cannot be guaranteed.     
5. The PI is billed the month that the purification is performed.

Please note that a set of guidelines for BAC recombineering of transgenes and targeting vectors can be found at vanderbiltresearch.org in the TMESCSR protocol collection under "BAC recombineering and transgenesis."