Self Service Real Time PCR

Equipment
  • Two Life Technologies QuantStudio 12K Flex instruments are available for use by appointment.  The instruments are identified by the computer attached to them.  
  • The “desktop” instrument accommodates:
    • Standard 96 well (200 ul) plates
    • 384 well plates
    • Taqman Low Density Arrays (TLDA) microfluidic cards
  • The “laptop” instrument accommodates:
    • Standard 96 well (200 ul) plates
    • 384 well plates
  • We do NOT have the 96 well (100 ul) “fast” block.
  • The 12K Flex machines can run in fast mode using the standard plate with fast master mix, reducing your run time, if your assay has been optimized for fast mode. 
  • This equipment may be used for SNP genotyping, gene expression, CNV analysis and microRNA assays.
Location
  • Blalock 1003
  • To access this room before 9 AM, please knock on the door of Blalock 1005.  After 9 AM, check in with the staff in Blalock 1004.
Reservations
  • Scheduled on a first come, first served, basis using the iLab scheduling module.
    • Detailed instructions for using iLab are available. You must be a registered user and attached to your PI’s account in order to use the scheduling module. Everything your PI or financial administrator needs to know about setting up a lab in iLab is on the Core-in-a-box site.
    • There will be a $40 fee applied to all customers who extend past their reservation time, including the time necessary to retrieve data.  You must be done by the time your reservation ends.
    • There is an automatic 30 minute buffer between reservations.  You will not be charged for this buffer.
  • Investigators may reserve the machines by the hour, for assays recording real-time metrics, or in 15 minute time increments for end-point reads. 

Reserve A Time

Usage Instructions
  • All reactions should be set up by the investigators in their own labs and the plate brought at the appropriate step in the process for loading on the machine. 
  • The specialized centrifuge and staking equipment necessary for loading the microfluidic [TLDA] cards is available for use in our lab. 
  • At the completion of the run, investigators should return and retrieve their data on a flash drive. 
  • A typical gene expression run requires a 2.5-hour reservation to ensure that you have enough time to fill out run details and retrieve data. 
  • A fast run typically requires a 1.5 hour reservation
  • SOPs for inexperienced users are available.
Consumables:
  • Consumables for the runs may be ordered through the Core Store.
    • MicroAmp® Optical Adhesive Film (catalog 4311971) or the equivalent
    • MicroAmp® Optical 384-Well Reaction Plate (catalog 4343370 or 4309849) or the equivalent
    • Applied Biosystems Array Card (custom or off the shelf)
    • MicroAmp® Optical 96- Well Reaction Plate (N8010560) or the equivalent
    • Applied Biosystems TaqMan Reagents or the equivalent
    • Applied Biosystems SYBR Green Reagents or the equivalent
  • Taqman Gene Expression, MicroRNA, Genotyping and CNV assays may be ordered through our Life Technologies ordering tool. 
  • Generic assay primers may be ordered through any of our oligonucleotide ordering portals.
Notes for users:
  • Please arrive on time for your time slot. It is best to arrive about 5 minutes early. If you start your run late and it runs past the time you have reserved, you will be charged $40 plus the additional time.
  • Use only optical plates and lids that are designed for this instrument.  Use of other plates and covers may adversely affect your data and our equipment.
  • Please do not write with marker on the top of the plate. Write ONLY on the sides of the plates, otherwise the marker will contaminate the machine.
  • If you reserve a time slot and don’t call to cancel it at least an hour prior to use, you will be charged for the time slot.
  • If you are using the 6 AM slot, you can set up your reactions the previous evening, store them wrapped in foil at 4 degrees overnight and bring them in the morning.
  • Data should be analyzed using Expression Suite from Life Technologies.  There is also a cloud based version of this software. 
  • Our instruments are typically calibrated to quantitate FAM, VIC & SYBR (or their analogs).  The instruments are capable of visualizing up to three other dyes, however, so if you have a specialized experiment, please contact us to discuss it.
  • If you are new to qPCR and would like to learn more about it, ThermoFisher has a great YouTube Channel with answers to many questions.
Pricing:
  • $20 per hour (internal JHU customers)
  • $10 for a 15 minute end point read
  • $40 overtime usage fee in addition to actual time used (this includes time spent collecting data).  You must be done in the lab by the end of your reserved time.

Reserve A Time