- Quantitative PCR facility
- Optimized HIV-1, SIV and cellular (housekeeping) primer/probe sets, Nucleic acid standards & other reagents
- Consultation & training in primer/probe design, data analysis, etc.
- Viral genetic analysis (from viral RNA or proviral DNA)
- Custom Molecular Support
Core Service Contact:
Farida Shaheen, PhD.
Technical Director, Viral and Molecular Core
Quantitative PCR Facility
The Core recently acquired an ABI 7500FAST realtime quantitative PCR machine (located in Room 515 of the Johnson Pavilion). Using dual labeled probes (Molecular Beacon & Taqman Technology),
the system is capable of reliably detecting two-fold differences in DNA
or RNA template concentrations over a range of 10-1,000,000 template
copies. The machine is available for use to all Penn CFAR members,
members of Department of Medicine, Neurology, and others (please inquire).
This machine has several new and valuable features:
- Run time is 1 hour with FAST mode (1:50 hour for standard mode & 2:30 hr for SYBR Green with melting curve)
- Compatability with 10 - 30 ul volume reactions (with considerable savings of reagents)
- Capability to view amplification plot develop in realtime
- Improved data analysis capability with simultaneous standard curve development for multiple targets
To maintain smooth operation, a monthly calendar is attached to the machine for researchers to sign up and reserve run time in advance. An additional sign-up sheet is also attached to the machine for users to fill in information at the time they are actually using the machine.
To assist investigators with data analysis custom primer/probe design for individual targets of interest, we have a workstation available with Sequence-Detection Software (data analysis) and Primer-Express Software (custom primer/probe sets) located in room 520 of Johnson Pavilion. Training in the use of this software is available through the core.
The charges for use of real time PCR machine is: $27.50 per 7500 Fast mode run ($37.50 per run for 2 hr or longer time run). Access to the Primer Express workstation is without charge.