Real Time PCR

Self-Service Real Time PCR Equipment 

Two Life Technologies QuantStudio 12K Flex instruments are available for use by appointment. 

The instruments are named after our favorite Muppet scientists, Dr. Bunsen Honeydew and Beaker. 

Instrument #1
(Dr. Bunsen Honeydew)
  • Standard 96 well (200 ul) plates
  • 384 well plates
  • Taqman Low Density Arrays (TLDA) microfluidic cards
Instrument #2
(Beaker)
  • Standard 96 well (200 ul) plates
  • 384 well plates

Please note: 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.

How to Reserve

All equipment reservations are scheduled on a first come, first served, basis using the iLab scheduling module.

reserve time
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.

How it works:

  • Investigators may reserve the machines by the hour, for assays recording real-time metrics, or in 15 minute time increments for end-point reads. 
  • 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.

Equipment Location

Johns Hopkins East Baltimore Medical Campus

600 N. Wolfe Street

Blalock Building, Room 1003

Baltimore, MD 21287 USA

NOTE: 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.

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.

Policies

  • 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.

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 equivalentApplied 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.


click here for core store ordering

New to iLab? Don’t have an account?

All reservations are processed through the JHU Agilent iLab system. Prior registration is required before equipment can be used.

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.

Account Registration

To register for an account:

1. Open a web browser and go to the JHU iLab page. Click on the blue “Sign up” button in the upper right hand corner.

2. Login using your Johns Hopkins University credentials in the section labeled “JHU Users”. If you are not a Johns Hopkins University user, click the prompt in the “Not a Johns Hopkins University User” box.

3. Upon successful authentication, if you are a Johns Hopkins University user you will be prompted to select your PI/lab from the pull-down list and provide your phone number. If you are a not a Johns Hopkins user, iLabs will send you a welcome email and login credentials. Submit the completed registration form.

4. You will be sent an email with basic instructions on using the system. Please note that lab members will not be able to place orders with the new account until a PI has approved the lab membership request and assigned a budget number.