University of Pennsylvania
DNA Sequencing Facility
Large Sequencing Projects
Description of Service
The facility will sequence through DNA clones by primer walking or, if needed, deletion subcloning. Please fill out a Request for Sequencing Project Management form when submitting the sample, indicating if you wish to design primers yourself or delegate the responsibility to facility staff. Charges for primer walks are calculated according to the number of lanes sequenced and primers constructed. A clean-up treatment of sufficient template for the whole project (nominally 4-5 μg per kb for each direction) is recommended to produce quality sequence and maximize step size.
After a good quality template is obtained, each step (~500 nucl.) requires around one week, including primer design and synthesis, sequencing, sequence evaluation, and assembly. Charges for deletion subcloning should be discussed with facility staff. Assembly of the overall sequence, ORF analysis, and homology searches can be performed, at no charge, if requested by the user.
The DNA Sequencing Facility is an Abramson Cancer Center Shared Resource that is approved and partially funded by the National Cancer Institute.