Real-time quantitative PCR (qPCR) analysis

TATAA Genomics Core Facility offers you a broad selection of real-time PCR instruments, including fast cyclers, High Resolution Melt (HRM) cyclers, and high throughput instruments. Applications such as gene expression analysis, genotyping, SNP analysis, HRM, digital PCR, Immuno-qPCR, and any more esoteric application of your choice are easily performed with confidence. TATAA’s experts can also advice on how to select the optimum reference genes or perhaps use alternative means to normalize your data.

Examples of real-time PCR analyses that can be performed at TATAA Genomics Core Facility are:

  • Gene expression analysis (relative and absolute quantification)
  • Genotyping
  • Copy number variation (CNV) analysis
  • miRNA analysis
  • DNA methylation analysis
  • CpG-methylation analysis
  • SNP typing
  • Allelic discrimination (end-point analysis)
  • Digital PCR
  • Protein quantification with immuno qPCR
  • qPCR data analysis and data mining with access to user friendly professional softwares

or other applications of your choice

 Our Göteborg facility’s real-time PCR instrument park is the most advanced in Europe. We have instruments from all leading manufacturers, several of which are capable of High Resolution Melt (HRM) analysis. You can pipette and setup reactions manually or use our automated liquid handling systems to save time and standardize procedures.

For your convenience plastic consumables such as plates, strips, tubes, pipette tips etc. for your experiments are included in our services. All have been tested for best performance by our experts, and all instruments are regularly calibrated with those consumables to ascertain optimum performance and highest possible repeatability and reproducibility. We are also carefully selecting real-time PCR reagents for optimum performance and lowest cost, available for you to purchase. For work in other labs, you can purchase the consumables from us separately to ascertain you can use the same protocols and stick to the same SOPs. Please visit Products for more information.

IN OUR FACILITY IN GÖTEBORG:


Pre-PCR lab

The Pre-PCR lab is a DNA-free zone. Here primers and probes are diluted and mastermixes are prepared. The preparations are made in a DNA-free hood, kept clean using UV-light. In the Pre-PCR lab there is also a scale to weigh e.g. chemicals and components of buffers etc.

PCR lab

In the PCR lab all PCR reactions are set up. Samples are diluted, templates and mastermixes are mixed and transferred to plates, strips, tubes etc. This is the lab where normalization, cDNA synthesis and real-time PCR is performed. In addition to several automated liquid handling systems and the real-time PCR instruments, general equipment include centrifuges, vortexes and a heatsealer.


Instruments – Real-time quantitative PCR (qPCR) analysis


Centrifuge

Eppendorf centrifuge 5804
The bench-top centrifuge 5804 from Eppendorf is a multi-purpose centrifuge feature a powerful, main-tenance free motor. A variety of rotors for different applications are available. It is possible to centrifuge plates for the qPCR applications as well as high volume centrifugation of Falcon® tubes just by changing the rotor. The rotor spins up to 14 000 rpm, 20 800 x g.


Liquid handling

Corbett Robotics CAS-1200
CAS-1200 pipetting station allows you to rapidly and reliably perform your pipetting automatically, preventing human mistakes. The station uses a single-channel pipette to automate PCR setup, e.g. distribute mastermixes and add samples. Up to 96 PCR reactions can be set up in approximately 30 minutes.

The instrument worktable is set up with a Master Mix Block, a Reagent Block, tips, and plates/tubes. Conductive tips enable high-precision pipetting with liquid level sensing. The software, which is operated on a PC next to the robot, is easy to learn and use. During the run the position of the pipetting head is highlighted on the screen as it moves, allowing the progress of a run to be followed in real-time. A feature that allows samples concentrations to be imported to the software directly from a text file is excellent when normalizing RNA or DNA concentrations. Other helpful features can be used when performing serial dilutions with variable dilution ratios or optimizing assays.



Eppendorf epMotion 5070
The Eppendorf epMotion 5070 is an automated pipetting system which enables fast and reproducible liquid handling by pipetting and dispensing. Pipetting protocols previously carried out manually can easily and quickly be transferred to the epMotion 5070 workstation. The workstation is programmed using an integrated control panel. By prompting the user to carry out necessary actions, programs are easy to follow and understand. The epMotion 5070 workstation enable quick filling of plates e.g. in the 384-well format. Optical sensors control and ensure the reliability of the procedure.


Quantitative real-time PCR instruments

IntelliQube®Instrument_IntelliQube

Fully Automated PCR Setup, Amplification and Analysis System.

The IntelliQube is a fully automated and high-throughput instrument featuring seamless integration for liquid handling, thermal cycling, detection and data analysis. IntelliQube leverages best-in-class multi-functional dispensing for high speed liquid handling in 768-well Array Tape®. Precise sample dispensing into Array Tape is handled by an interchangeable CyBi® FeliX 96- or 384-channel pipetting head using only 0.8 µl sample per reaction. Reagent dispensing is performed by the Douglas Scientific Dispense Jet, a 4-channel, non-contact dispense head that loads the assay mixtures required for your protocols using only 0.8 µl per reaction.

With CVs less than 5%, the liquid handling on the IntelliQube offers exceptional reproducibility and data quality. The IntelliQube supports inline amplification with a Peltier thermal cycling block designed specifically for Array Tape®.




Applied Biosystems QuantStudio™ 12K Flex Real-Time PCR System

An all-in-one qPCR instrument that is designed for maximum throughput, flexibility and scalability. With five interchangable thermal cycling blocks (33 nL – 100 µl reactions), you can analyze from 1 to over 12,000 data points per run in the assay format that is right for your project. Comprehensive software analysis tools for gene expression, genotyping and digital PCR are available.

TaqMan® Assays are available for all five thermal cycling blocks: OpenArray®, TaqMan® Array Card, 384-well and standard or fast 96-well blocks. With the QuantStudio™ 12K Flex OpenArray® Plate format enables testing from 48 samples against 18 unique assays to 12 samples against 224 unique assays. A streamlined workflow with the QuantStudio™ 12K Flex AccuFill™ System minimizes start-up and hands-on times. Data from many OpenArray Plates may be combined into the same project. With the TaqMan® Array Card it is possible to order plates with preloaded assays and just add sample and mastermix before run. The QuantStudio™ 12K Flex Real-Time PCR System has six decoupled excitation and emission filter channels for the greatest number of dye combinations and maximum multiplexing capabilities. It has four decoupled excitation and emission filter channels for the OpenArray® plates.

With the QuantStudio™ 12K Flex Real-Time PCR System TATAA Biocenter offers Digital PCR services, as well as High-throughput real-time quantitative PCR (qPCR) analysis services.



 

Bio-Rad CFX96 and CFX384 Real-Time System

The CFX96 optical reaction module converts a C1000 thermal cycler into a powerful and precise real-time PCR detection system. This six-channel (five channels for multiplexing) real-time PCR system combines advanced optical technology with precise thermal control to deliver sensitive and reliable detection. The CFX384 is built in a similar way but with five channels (four channels for multiplexing). The instruments features factory calibration to minimize setup time and thermal gradient to optimize reactions in a single run. Its optimized design lets you configure the CFX96 and CFX384 systems to fit your laboratory needs – run without a computer, run up to 4 systems from 1 computer, or integrate with the CFX automation system for higher throughput.




Fluidigm BioMark™ SystemInstruments_Fluidigm_BioMark HD

Analysis on the BioMark™ is only offered as a service. For more information about TATAA’s service offers, please visit
» Services – qPCR Analysis

The BioMark™ system is a high-throughput qPCR instrument that uses integrated fluidic circuits known as dynamic and digital arrays for the applications Digital PCR, Gene Expression, and Genotyping. The dynamic arrays are suited for either 48 samples and 48 assays (2304 reactions) or 96 samples and 96 assays (9216 reactions). The digital arrays has 12.765 chips (for 12 samples and 765 reactions/sample) and 48.770 chips (for 48 samples and 770 reactions/sample).

This innovative instrument allows practice of tried-and-true techniques, such as TaqMan® assays, while realizing a previously unachievable throughput. Researchers save on reagents, pipette tips and time. Far more data are generated compared to microwell chips, and with no pre-spotting required.

Use as little as 10 picograms of starting material – reproducible results from populations of cells down to a single cell.




Illumina Eco Real-Time PCR system 
The Eco system delivers everything a full-featured qPCR instrument offers and more. It has a proprietary thermal system for unrivaled temperature control and a sensitive optical system to facilitate all four-color multiplex applications.

48-well block
5-20 µl reaction volume
Has software for HRM




Roche LightCycler 480 Real-Time PCR System
The LightCycler ® 480 Real-Time PCR System is a high throughput gene quantification or genotyping real-time PCR platform. With exchangeable blocks for 96 and 384 samples in multiwell plates the instrument is nice to use when handling small amounts of samples as well as large amounts. The LightCycler ® 480 Real-Time PCR System can complete a 348 well PCR run in 40 minutes. The system is operated by a separate desktop data station based on Windows XP. The intuitive software includes step saving short cuts for high-throughput sample and data handling. One-click experiment set up with options to refine results later. The platform gives options for basic and advanced gene expression analysis. Work in a broad dynamic range from 1010 to a single copy in a single run is possible. Examples of projects to perform on the platform are genetic variation studies based on hybridisation probes, simple probes or hydrolysis probes. LightCycler®480 is highly robust and convenient when working with mutation discovery. The features for High Resolution Melting is useful for reducing time and costs compared to non- homogenous mutation screening methods that require PCR and analysis on separate instruments. The platform provides an outstanding specificity and sensitivity with high sample throughput.




Roche LightCycler Nano

The LighCycler Nano is a small qPCR instrument with excellent thermal precision and robust solid-state optics that provides complete spectral information. The instrument has 32 wells and uses standard 8-tube strips. The software is easy to use and different methods for quantification and mutation analysis are available as well as HRM analysis.



Stratagene Mx3005P
The Mx3005P software is easy to use due to the intuitive interface, quick experiment design, powerful data analysis and easy report generation. Due to the flexibility the systems are ideal for a wide variety of applications and chemistries. E.g. Gene Expression Analysis, Microarray Data Validation, SNP Genotyping, and Pathogen Detection.


Thermo Scientific PikoReal Real-Time PCR System

The PikoReal Real-Time PCR System is a highly flexible gene quantification and genotyping platform in a 96-well block format. It incorporates innovative, energy-saving technologies to achieve fast performance while meeting the highest thermal requirements – all with reduced energy, plastics, and reagent consumption for real cost savings.

It uses Ultra Thin Wall (UTW) Piko PCR plates that are 25% the size of conventional plates, yet compatible with multi-channel pipettes, reagent dispensers, and automated liquid handling systems. Five detection channels enable up to four target multiplexing and it has a high temperature uniformity across the wells throughout the temperature range. The PikoReal Real-Time PCR System has an easy-to-use software and offers users unlimited access to the PikoReal Software, which introduces the Virtual Pipetting Tool for conveniently creating and editing the plate layouts. The PikoReal software offers extensive analysis options within multiple analysis modules.


 


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