Self Service Real Time PCR
- Two Life Technologies QuantStudio 12K Flex instruments are available for use by appointment. Both instruments accommodate:
- Standard 96 well (200 ul) plates
- 384 well plates
- Taqman Low Density Arrays (TLDA) microfluidic cards
- 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.
- Blalock 1003
- **During COVID-19 Dedensification, please knock on Blalock 1005 and someone will let you into the room. Please maintain 6 feet separation during this process.
- Scheduled on a first come, first served, basis using the iLab scheduling module.
- **COVID-19 Dedensification Restrictions:
- Two time slots: 6AM-9AM or 10AM-1PM
- One user per time slot
- If you would like to run more than one plate, reserve the early time slot and email us and ask us to extend your reservation.
- There will be a $20 fee for all canceled reservations
- 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.
Reservations for independent use are available from 7 AM through 4 PM each day. Customers requiring assistance may make reservations from 9 AM -2 PM each day. The 4 PM slot runs overnight.
- Investigators may reserve the machines by the hour, for assays recording real-time metrics or in 15 minute time increments for end-point reads.
- 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 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
- 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 there is a run scheduled after yours, we will be forced to interrupt your run to start the next customer’s run.
- 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 that can handle larger sample sizes.
- 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.
- $20 per hour (internal JHU customers)
- $10 for a 15 minute end point read
- $20 flat cancelation fee
- $40 overtime usage fee (this includes time spent collecting data). You must be done in the lab by the end of your reserved time.