North Florida/South Georgia Veterans Health System
Gene Expression Core Laboratory
The Gene Expression Core Laboratory (GECL) provides the Gainesville VA Medical Center (GVAMC) and the University of Florida investigators with a state-of-the-art facility to define/compare gene expression patterns. To date, these include the ABI 7500 System for quantitative real-time polymerase chain reaction (qRT-PCR) analysis, and the Digital Imaging System, KODAK Image Station 4000MM. In addition, the core serves the GVAMC community by providing a resource for training, education, and consultation in modern gene expression and proteomic techniques.
The Kodak Image Station allows Luminescence, Multi-wavelength Fluorescence, Absorbance, Radioisotopic, and X-Ray detection with high resolution. The applications include images of stained, fluorescent, and P 32 labeled gels and other materials, blot membranes, and X-Ray detection with simultaneous fluorescent imaging.
The 7500 Real-Time PCR System monitors the PCR reaction at each cycle of the run and estimates the log phase cycles. It compares the curve generated to that of a known standard and calculates a starting copy number. The system consists of a 96-well thermal cycler with a built in fluorimeter. The software packages for quantitation and primer design are installed on an IBM compatible PC.
The core lab provides PCR plates and caps at a discount fee. Users are responsible for purchasing their own kits and other consumables. The core maintains the system and offers directions on protocols, and training sessions. There are a large number of protocols that employ Taqman chemistries. These include DNA quantitation and qRT-PCR using a dual-labeled sequence-specific probe, which is cleaved by the exonuclease activity of the TAQ polymerase and detected by the fluorometer. Other methods employ SybrGreen, which intercalates with double stranded DNA and is detected as the product is generated. There is also a protocol for allelic detection, which is used for identifying known mutations or polymorphisms. Assays for a number of genes have been developed, along with a new Assays-by-Design program available from Applied Biosystems.
Submission Policy & Requirements:
User accounts will be created upon request. To register, please contact Dr. Elena Yarmola, VA extension 4305 and receive initial instructions.
PLEASE NOTE: Each user must use his/her personal User ID and password to sign in to the 7500 RT-PCR system or Kodak Image Station. Users are required not to share the user ID and password with anybody. Please log off before allowing the next person to use the system, or shut down the instrument and computer if nobody else is waiting for the run. Failure to comply with the user policy will cause blocking or deleting the user account. The Principal investigator is responsible for verifying all information, for the PCR system use charges, and carries financial responsibility for the damage to the instrument if caused by misuse and/or neglecting the rules by his/her employees.
The plates and caps can be purchased from Applied Biosystems or from the core lab at a discount fee. For VA-funded investigators who pays charges from VA account the charges for the system use will be calculated as follows. Day runs: $12.00 per hour. Overnight runs: $30 for the first three hours plus $1 per every additional hour. The runs that started before 4 pm are considered as day runs, and the runs that started after 4 pm and finished on or before 10 am next day are considered the overnight runs. The run time between 10 am and 4 pm is considered as a day run time and is charged $12 per hour. For the investigators who pay charges from a non-VA account, all charges are 50% higher. Data analysis only (computer program without the PCR instrument use is free of charge).
After each run, please shut down the instrument and computer and write your name, date, and time of the run in the PCR log book. If the next person is going to use the instrument right after you, please make sure you log off from the computer before you let the next person use it, since the time before you log off is considered as your run time.
The charges for the use of Kodak Image Station for non-VA investigators are $30 per hour or $0.50 per minute of the Instrument use. Data analysis only (computer program without the PCR instrument use is free of charge). For VA investigators, instrument use is currently free of charge although charges may be introduced in future upon decision of Equipment Committee. After each use, please log off or shut down the computer and write your name and date in the Kodak log book. Please note that VA Radiation Safety Training is required to work with radioactivity.
If research supported by the GECL-GVAMC results in publication, please acknowledge this support by including the following in your publication(s): We would like to thank the North Florida/South Georgia Veterans Health System and the Department of Medicine at University of Florida School of Medicine in Gainesville, Florida, for the use of the Gene Expression Core Laboratory which provided __________ service.
Application Services Form:
The Application Service form can be obtained from Dr. Elena Yarmola, VA extension 4305. Please file the form electronically, print and obtain needed signatures.
In particular, please note that e-mail communication may not be secure against interception across the Internet. Please do not include sensitive information such as medical records or financial information, drug prescriptions, or your social security number.