Self Service Real Time PCR
Two Life Technologies QuantStudio 12K Flex machines are available for use by appointment, in Blalock 1005. All instruments accommodate standard 96 (200 ul) & 384 well plates as well as Taqman Low Density array (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.
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 3-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
Reservations are scheduled on a first come, first served, basis using the iLab scheduling module. Detailed instructions 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. 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.
Consumables for the runs may be ordered through the Core Store. Please see our list of recommended consumables for part numbers. 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.
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.
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 7 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. The 12K Flex machines generate .eds files, the 7900 generates .sds files. Both import into Expression Suite.
- $20 per hour (internal JHU customers)
- $10 for a 15 minute end point read