http://www.appliedbiosystems.com/

The Gene Expression Resource currently houses two ABI 7900HT Sequence Detection Systems. The ABI Prism 7900 HT Systems use Peltier-based thermal cycling, TaqMan chemistry, and extended-life 488 nm argon-ion laser excitation to detect and quantitate nucleic acid sequences. The systems can be used for gene expression, quantitation, and SNP detection using fluorogenic 5’ nuclease assay.

The primary use of this system is to examine relatively few numbers of genes across a wide spectrum of conditions.  This technology is very sensitive in that it is able to detect single copies of genes and requires very little starting material.  The system monitors PCR at each cycle of a reaction, and estimates the cycle when the reaction reaches log phase increments (when the most useful quantitative information about the sample is available). Quantitation can be "relative" to an internal standard such as a housekeeping gene, or "absolute" when compared to a standard curve generated from known concentrations. 

Quantitative Real Time-PCR can be performed using dual-labeled sequence-specific probes (ABI's Taqman probes), fluorogenic light-upon-extension primers (Invitrogen's LUX primers), or SYBR-based primers. For the most up-to-date information, tutorials, and manuals, please visit a few of our favorite sites: Applied Biosystems QRT-PCR , Invitrogen QRT-PCR, and LUX Primers.

 For allelic detection and SNP detection, ABI has a variety of pre-developed assays for specific genes. For a list of these assays and other protocols visit ABI DNA Seq. 

To learn more about RT-PCR, view webinar.

Usage / Requirements

Those interested in utilizing any of the Resource’s equipment must first speak with Jenniffer Rothschild, ext. 3958. She will arrange a time for you to be trained on the equipment, and to set up an account so that you may schedule equipment time. Once an account has been established all equipment scheduling is done via Resource Scheduling in Outlook Express Public Folders.

The investigator is responsible for purchasing kits and consumables. The Resource will maintain the instruments, provide training, and make primer design and analysis software available. Primer Express v2.0 software is available through the facility, call x1740 or x3958 for more information. There is a $25.00 charge for each plate run (up to 3 hours).

Please note the following criteria for the ABI 7900 Systems:

• The only sample containers that can be used are Semi- or Half-Skirted well plates with flat-tops. Users can not use strip-tubes, rounded-caps, or unskirted plates. Users will also need to purchase flat-top, optical caps or 0.2 mm thermal microseal film for their plates. All other plates and caps will not work with these machines. These items may be purchased through any company (ex: VWR or Krackeler.)

• Microsoft Windows platform is used instead of Macintosh.

• Machine 1 is calibrated for a 96-well plate.  Machine 2 is calibrated for a 96- and 384-well plates.  At this time, we do not own a block for Microfluidics cards, but if you are interested, we could get one. Just let us know.  

 JL 100207