The DNA Core Facility currently has an AB 7900HT real-time PCR machine available for use.  The 7900HT is a real-time PCR machine that utilizes LASER based fluorescence detection and PC software.  To find out more information on the 7900HT please visit the Applied Biosystems website at www.appliedbiosystems.com.

1. To get started on the real-time machine please read the following documents:

Essential of Real-Time PCR
Guide to Relative Quantitation
Relative Quantitation UB#2
QiagenRT-PCR

2. Next contact Angie Traner at 475-7844 and set up a time to discuss protocols and help on getting started.

3. Order your primers and probes. Isolate your RNA. If you are doing onestep, make cDNA.

4. Just before your first run you need to stop by the Core Facility for a brief overview of the machine and our policies and procedures and sign a usage agreement before you will be able to sign up for time on the machine..  Time is reserved by going to: coreweb.icmb.utexas.edu(http://146.6.133.14/) and either login with you DNA sequencing account or create a new account and log on to the "coreCalendar" and request access to the calendar.

5. Sign up for time for your first run. You should contact Angie Traner at 475-7844 and make sure she has time to help on your first run.

6. Set up your reactions. Come to MBB 1.426P, sign on the 7900HT and begin your first run.