Section of Research Resources:

Core Facilities

Quantitative Real-Time PCR

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IMMEDIATE UPDATE

7900HT and the 7300 are both available.

The SDS2.2.2 software is on a computer in C1734B. Some plates and covers are there, too. We'll keep it stocked.

System software for the 7300 is also installed on the computer.

You sign on with your e-pass ( or the person currently signed on lets you create your file ).

You will setup the plate as if you are running it on the instrument ( and type the barcode into the initial window that you would scan into ). The detector list has been imported.

(NOTE: If you add a new detector, you will need to export that so it can be uploaded onto instrument computer. Call Rob (x5823) ( or e-mail ) for instructions.)

Save the SDS file you create in the network folder \\hersheymed\files\Research\CoreFacility\Results\Functional Genomics Incoming in the Incoming 7900 Plates folder. Save as -- YourName_PI_BARCODE

The plate is then put in the refrigerator in C1734 and we collect them once a day ( by 1:30 PM ).

The same protocol should be followed with the plates for the 7300. Please make sure to label the skirt with your name and PI information. If you are setting up multiple plates on the same day, make sure there is a unique identifier on each.

(NOTE: System software you will give a message that it "Cannot locate the instrument". Hit Cancel to continue to setup ).

It's basically .. create the file and save it on the network. We pull that file up and run the plate and put the file ( now with data ) back onto the network the next day for you to analyze ( again, on the computer in C1734B).

And if you miss the 1:30 deadline, the plate can wait until the next day and/or you can bring it over by 4:30 or so.

If you need assistance please contact Rob or Georgina to schedule a time that we can help.

Primer Design and Purchase:

Primary recommendation is to utilize pre-optimized Gene Expression Assays, available from Applied Biosystems as well as others, to save you on time and optimization of primer and probes and concentrate on your experiment.

A frequently updated list of validated primer/probe sets for quantitation of a variety of mRNAs is maintained at http://www.realtimeprimers.org/
To design your own primer/probe sets, you can use Primer Express, which is only available for installation on your PC computer (Contact Rob Brucklacher at extension 5823) (see also FYI other primer design programs on the WEB).
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- Assays on Demand for Gene Expression, SNP Analysis, Genotyping (From ABI):
  We recommend ABI's Assays on Demand (Click here to search for an assay) for most uses when available, as the expense of purchasing these pre-validated assays is far out-weighed by the savings in time necessary to test and validate newly designed assay primers.
- Please check with us for current pricing of an Assay on Demand or Assay by Design primer set..
  
  Note: Please consult with us before ordering your probes to ensure that our equipment is compatible with the selected dye.

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Pricing for reagents, supplies and service:

To be updated as soon as possible

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For further information on Quantitative Real Time PCR analyses, please contact Rob Brucklacher or Georgina Bixler at extension 5823, Hershey Center for Applied Research, Room 3200

Page maintained by Bruce A. Stanley
Last revised: July 05, 2007

Penn State Milton S. Hershey Medical Center c2001
This page was last updated on January 29, 2003
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