Direct sequencing of BACs, PACs, P1s, and difficult Cosmids is now available at the DNA Analysis Facility, due to advances in Dye Terminator chemistry, the addition of Thermofidelase I, and an extended thermal cycling protocol developed by Bruce Roe et al. at Oklahoma University. The price for BAC sequencing is $25 due to increased reagent usage and a larger purification column.
To place a sequencing request for BAC, PAC, P1, or Cosmid DNA to be run using the BAC protocol, you must choose BAC, PAC, P1, or difficult Cosmid when selecting the type of template. This will ensure that we use the correct protocol.
Please Note: Most cosmids can be sequenced directly using our standard protocol. If your cosmid has an unusually large insert, or if a previous attempt at direct sequencing has failed, the BAC protocol is recommended.
Template and primer must be premixed in a single tube at the following concentrations and volumes:
| Template concentration: | 0.5 µg/µl |
| Primer concentration: | 4 µM |
| Template volume: | 4 µl |
| Primer volume: | 5 µl |
| TOTAL VOLUME: | 9µl |
Please note that due to the nature of BAC sequencing, turnaround times vary.