Automated qPCR Setup with the CyBio Felix Liquid Handler
Quantitative PCR (qPCR), also known as real-time PCR, is a core technique in modern molecular biology and diagnostics due to its high sensitivity and specificity in detecting nucleic acids. However, setting up qPCR reactions manually can be a tedious process, especially with large numbers of samples. Labs all over the world are implementing automated qPCR setup to improve throughput and reduce hands-on time while conserving reagents and improving the quality of the results.
A robotic liquid handler is at the core of an automated qPCR setup workflow. The CyBio Felix is a robust liquid handling platform that can pipet small volumes with high accuracy and precision - essential for highly reliable qPCR results.
Benefits of automating qPCR setup processes with CyBio Felix
Among the many advantages of working with a CyBio Felix automated liquid handling platform, these key features are particularly beneficial for automating qPCR setup:
- Pipetting flexibility – the CyBio Felix platform can handle a range of volumes and pipetting formats (single channel, rows, column, full plate) within a single run as well as perform both forward and reverse pipetting to accurately mix and dispense reagents of different viscosities.
- Versatility in small footprint – the dual deck design provides 12 usable deck positions within a footprint much more compact than other platforms, so it can meet the challenge of automating your workflow.
- High precision and accuracy – thanks to Analytik Jena’s CyBio pipet sealing technology and well-proven pipetting heads and adapters, the CyBio Felix reliably delivers the correct volumes from 1.0 to 1000 µL.
- Ease-of-use for both operators and protocol developers- Application Studio software allows laboratory staff to operate CyBio Felix easily by selecting approved protocols and following setup instructions. Developers can access CyBio Composer’s scripting environment, which allows creation of optimized protocols with configurable parameters.
Workflow for automated qPCR setup with CyBio Felix (standard throughput)
Deck setup
Place an empty microcentrifuge tube as well as tubes with the following reagents: buffer, Taq polymerase, dNTPs, PCR-grade-H2O, forward and reverse primers in a passive cool block for 2mL tubes.
Next, place the cooling block, an empty qPCR reaction microplate, and the plate containing the DNA template on to the deck of the Felix. Add tip boxes and racks as necessary.
Automated master mix preparation
Using a fresh disposable tip for each reagent, Felix will add the necessary volumes to the empty tube to create the qPCR master mix.
With a volume range of 1.0 µL to 1000 µL, the CyBio FeliX Choice pipetting head can handle any of your PCR reagents. By automating master mix preparation, you can generate identical master mixes day after day, to help ensure data reproducibility.
FeliX tip: Let the FeliX software tell you how much reagent you need for each run!
Automated dilution of DNA samples
If necessary, Felix can perform automated sample dilution to achieve DNA concentrations within the quantifiable range of the qPCR reaction.
Transfer of master mix to qPCR plate
Using a single tip, Felix will transfer the correct amount of qPCR master mix from the microcentrifuge tube to each well of the reaction plate.
FeliX tip: Aspirate a large volume and reverse pipette your master mix for quick and precise setup!
Transfer of samples to the qPCR plate
CyBio FeliX can accurately transfer as little as 1.0 µl of your sample template to the reaction plate, using single-, eight-, twelve- or 96/384- channel pipetting format that best fits the layout and number of samples you need to process.
FeliX tip: Let FeliX pre-mix your samples to eliminate the need of vortexing and centrifuging the reaction plate.
Run the qPCR analysis
Remove the plate from the CyBio Felix deck, seal the plate, then place it in a qPCR instrument such as one of our qTOWER iris real-time thermal cyclers.
Workflow for high-throughput qPCR setup
If your laboratory routinely processes more than 96 samples for qPCR analysis, the following options can reduce the time required for automated qPCR setup:
- Prepare master mix in one row or column of a deep well microplate or in a reservoir for multi-tip access.
- Use an 8/12 channel adapter to dispense master mix to an entire column/row of the reaction plate at a time.
- Use an 96/384 channel pipetting head to transfer and mix DNA samples with qPCR master mix.
- Integrate the CyBio Felix with a qTOWER3 Auto, plate sealer, and CyBio Carry for an end-to-end laboratory automation solution.
How to Equip your CyBio Felix for automated qPCR setup (standard throughput)
Hardware and accessories
CyBio FeliX Basic Unit with Enclosure
OL5015-24-100
Adapter 24, passive cooling function
Holds 24 tubes, 1.5 or 2mL. Pack of 2.
844-00136-0
Adapter 96, passive cooling function
844-00135-0
Compatible with semi-skirted and full-skirted 96-well microplates.
40 mm Height Adapter
844-00445-0
pack of 2
70 mm Support
OL3317-11-110
Waste Box & Bags
Waste Box for used tips:
844-00430-0
Waste bags for use with waste box
(pack of 50):
10-406-342
Consumables
CyBio TipBox 96/250 µL (Filter Tips)
OL3811-25-937-F
Optical sealing foil (77 x 140 mm)
846-050-258-5D
Additional materials required for automating qPCR setup
- qPCR reagents (Taq polymerase, buffer, dNTPs)
- primers
- sample DNA (template)
- microfuge tubes
Related topics and products
Application Notes – automated qPCR setup
Downloads
The newsletter of Analytik Jena frequently keeps you posted about:
- News
- Trends and developments
- Events