qPCR Machines and Real-time PCR Platforms

Features of Real-time PCR Platforms and qPCR Machines

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications
In weighing up the pros and cons of the different real-time PCR machines available for your laboratory, factors to consider include: supported chemistries; multiplex capability for that chemistry; throughput; flexibility; format; easy-of-use and robust software package; reproducibility; speed; size; technical support; customer support and not least the cost, not only of the initial equipment outlay and servicing but also the associated cost of consumables and reagents. It is also possible to 'try before you buy', most companies will provide a loan machine. It is wise to test a few of these once you have narrowed down your choice. User experiences should not be overlooked and there are a number of useful websites and forums where you can address you questions and queries.

Lab-on-a-Chip Technology (Vol. 2): Biomolecular Separation and Analysis

Edited by: Keith E. Herold and Avraham Rasooly
Applications in the biomedical and life sciences: biomolecule separation, electrophoresis, chromatography, protein and cell separation, genetic and transcriptome analysis, PCR, cell viability analysis and microorganism capturing.

"a comprehensive and felicitous compendium" (Drug Research) read more ...

The following PCR machines are compared for various features to help you decide which instrument is most suitable for your needs. Please scroll down the page to see the various platforms.

Applied Biosystems: ABI 7300, ABI 7500, ABI 7500 Fast, ABI 7900 Fast HT with automation accessory, ABI StepOne
Roche: LightCycler 480, LightCycler 1.5, LightCycler 2.0
Stratagene: Mx4000, Mx3000P, Mx3005P
Cepheid: SmartCycler
Corbett: Rotor-Gene 6000
Eppendorf: Mastercycler ep realplex
BioRad: MiniOpticon, MyiQ, Opticon2, Chromo4, iQ5

Company	Applied Biosystems	Applied Biosystems
Model	ABI 7300	ABI 7500
Laser/Lamp	tungsten halogen lamp	tungsten halogen lamp
Detector	CCD camera	CCD camera
Thermocycling	peltier element block	peltier element block
Excitation spectrum	single excitation filter	five excitation filters
Filters/detection channels	4-position filter wheel: FAM/SYBR Green I, VIC/JOE, NED/TAMRA and ROX	5-position filter wheel: FAM/SYBR Green I, VIC/JOE, NED/TAMRA/Cy3, ROX/Texas Red and Cy5 dyes
Format	96-well plates or 0.2 ml tubes	96-well plates or 0.2 ml tubes
Time (40 cycles)	1.75 hours	1.75 hours
Reaction Volume	20-100 μl	20-100 μl
Fluorescence Chemistry	hydrolysis probes and SYBR Green I, other chemistries possible but not supported	hydrolysis probes and SYBR Green I, other chemistries possible but not supported
Multiplexing Supported	4-plex	5-plex
Passive Reference	ROX (optional)	ROX (optional)
Dimensions (H x W x D)	34 x 49 x 45 cm	34 x 49 x 45 cm
Weight	29 Kg	34 Kg
Other features	primer/probe design software included	primer/probe design software included
	PCR mastermix kits, parameter specific kits	PCR mastermix kits and parameter specific kits
	Assay by Design and Assay on Demand Services	Assay by Design and Assay on Demand Services
	Software: gene expression analysis, absolute quantification, allelic discrimination and plus/minus assays	Software: absolute quantification, relative quantification, allelic discrimination, isothermal and plus/minus assays
		SDS v1.4 21CFRp11 Module assists with 21CFRp11 compliance

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	Applied Biosystems	Applied Biosystems
Model	ABI 7500 Fast	ABI 7900 Fast HT with automation accessory
Laser/Lamp	tungsten halogen lamp	argon ion laser
Detector	CCD camera	CCD camera
Thermocycling	peltier element block	peltier element 9700 block
Excitation spectrum	five excitation filters	488 nm
Filters/detection channels	5-position filter wheel: FAM/SYBR Green I, VIC/JOE, NED/TAMRA/Cy3, ROX/Texas Red and Cy5 dyes	500-660 nm continuous wavelength detection, SYBR Green I, FAM, VIC, JOE, NED, ROX, TAMRA, TET
Format	fast 96-well plates optimised for 10 μl reactions	96- and 384-well plates (interchangeable blocks)
Time (40 cycles)	< 30 mins	33 mins (96-well plate) or 52 mins (384-well plate)
Reaction Volume	10-30 μl	5-20 μl
Fluorescence Chemistry	hydrolysis probes and SYBR Green I, other chemistries possible but not supported	hydrolysis probes and SYBR Green I, other chemistries possible but not supported
Multiplexing Supported	5-plex	6-plex
Passive Reference	ROX (optional)	ROX (optional)
Dimensions (H x W x D)	34 x 49 x 45 cm	64 x 125 x 84 cm
Weight	34 Kg	114 Kg
Other features	primer/probe design software included	primer/probe design software included
	PCR mastermix kits, parameter specific kits	PCR mastermix kits, parameter specific kits
	Assay by Design and Assay on Demand Services	Assay by Design and Assay on Demand Services
	Software: absolute quantification, relative quantification, allelic discrimination, isothermal, melt curve analysis and plus/minus assays	Software: absolute quantitation, relative quantitation, allelic discrimination, isothermal, plus/minus assays, high throughput SNP genotyping and gene expression
	SDS v1.4 21CFRp11 Module assists with 21CFRp11 compliance	SDS v1.4 21CFRp11 Module assists with 21CFRp11 compliance
		Robotic plate un/loading for up to 84 plates, high throughput: real-time PCR >5000 wells per day and end-point 30,000 wells every 3 hours

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	Applied Biosystems	Roche
Model	ABI StepOne	LightCycler 480
Laser/Lamp	LED	high intensity xenon lamp
Detector	3 emission filters and photodiode	CCD camera
Thermocycling	peltier element block	peltier element block incorporating Therma-Base technology
Excitation spectrum	480 nm	430-630 nm, 5-position filter wheel: 450, 483, 523, 558 and 615 nm
Filters/detection channels	3 filters: SYBR Green I/FAM, VIC/JOE and ROX	6 position filter wheel: 500, 533, 568, 610, 640, 670 nm
Format	48-well plates, 0.1 ml individual or 8-strip tubes	96- and 384-well plates (interchangeable blocks)
Time (40 cycles)	< 40 mins	< 1 hour (96-well plate) or < 40 mins (384-well) plate
Reaction Volume	10-30 μl	10-100 μl (96-well plate), 3-20μl (384-well plate)
Fluorescence Chemistry	hydrolysis probes, SYBR Green I, other chemistries possible but not supported	hydrolysis probes, hybridisation probes, simple probes, SYBR Green I, UPL
Multiplexing Supported	3-plex	3-plex hybridisation and 6-plex hydrolysis probes
Passive Reference	ROX (optional)	Not required
Dimensions (H x W x D)	24.6 x 51.2 x 42.7 cm	54.5 x 60 x 60 cm
Weight	23.6 Kg	55 Kg
Other features	primer/probe design software included	primer/probe design software available
	Touchscreen, wizard driven software with networking capabilities	System compatible with 21 CFR part 11 requirements
	PCR mastermix kits	PCR mastermix kits, parameter specific kits
	Software: standard curve, relative standard curve, comparative Ct (DDCt), allelic discrimination, melt curve analysis and plus/minus assays	Software: absolute quantification, Tm calling, colour compensation, qualitative detection, statistical analysis, relative quantitation, high-resolution melting, genotyping, LIMS/bar-code module

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	Roche	Roche
Model	LightCycler 1.5	LightCycler 2.0
Laser/Lamp	LED	LED
Detector	3 photodetection diodes	6 photodetection diodes
Thermocycling	heated air	heated air
Excitation spectrum	470 +/- 10 nm	470 +/- 10 nm
Filters/detection channels	3 channels: 530, 640, 710 nm	6 channels: 530, 560, 610, 640, 670, 710 nm
Format	32 glass capillaries	32 glass capillaries
Time (40 cycles)	30 mins	30 mins
Reaction Volume	20 μl	20 μl or 100 μl
Fluorescence Chemistry	hybridisation probes, hydrolysis probes, molecular beacons, SYBR Green I	hybridisation probes, hydrolysis probes, molecular beacons, SYBR Green I, simple probes, UPL
Multiplexing Supported	2-plex hybridisation probes	4-plex hybridisation and 2-plex hydrolysis probes
Passive Reference	Not required	Not required
Dimensions (H x W x D)	45 x 30 x 40 cm	45 x 30 x 40 cm
Weight	20 Kg	20 Kg
Other features	primer/probe design software available	primer/probe design software included
	PCR mastermix kits, parameter specific kits	PCR mastermix kits, parameter specific kits
	Software: absolute quantitation, genotyping analysis, melting curve analysis, multiplex analysis	Software: absolute quantification, relative quantification, genotyping analysis, Tm calling analysis, multiplex analysis, plus/minus assays
	Relative quantification software available	Nucleic acid quantitation
		Conforms to European directive for in vitro diagnostic medical devices 98/79/EC.

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	Stratagene	Stratagene
Model	Mx4000	Mx3000P
Laser/Lamp	Quartz tungsten halogen lamp	Quartz tungsten halogen lamp
Detector	4 PMTs	1 scanning PMT
Thermocycling	Solid-state resistive/convective peltier hybrid block	Solid-state peltier-based block
Excitation spectrum	4 interchangeable filter wheels in the range 350-750 nm	4 user defined matched filter wheels in the range 350-750 nm
Filters/detection channels	4 interchangeable filter wheels in the range 350-830 nm	4 user defined matched filter wheels in range 350-700 nm (default: FAM, Hex, ROX, Cy 3)
Format	96-well plates, 0.2 ml tubes, 8 x 0.2ml strips	96-well plates, 0.2 ml tubes, 8 x 0.2ml strips
Time (40 cycles)	< 1.5 hours	< 1.5 hours
Reaction Volume	10-50 μl	10-50 μl
Fluorescence Chemistry	hydrolysis probes, fret probes, molecular beacons, Scorpions, Amplifluor, SYBR Green I	hydrolysis probes, fret probes, molecular beacons, Scorpions, Amplifluor, SYBR Green I
Multiplexing Supported	4-plex	4-plex
Passive Reference	Optional ROX	Optional ROX
Dimensions (H x W x D)	76 x 46 x 51cm	33 x 46 x 43 cm
Weight	50 Kg	20 Kg
Other features	PCR mastermix kits	PCR mastermix kits
	System compatible with 21 CFR part 11 requirements	System compatible with 21 CFR part 11 requirements
	Software: gene expression analysis, allelic discrimination, melting curve analysis	Software: gene expression analysis, allelic discrimination, melting curve analysis
	Integrated computer	Integrated computer

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	Stratagene	Cepheid
Model	Mx3005P	SmartCycler
Laser/Lamp	Quartz tungsten halogen lamp	4 high intensity LEDs
Detector	1 scanning PMT	Silicon photodetectors
Thermocycling	Solid-state peltier-based block	Resistive heating of ceramic plates with forced-air cooling
Excitation spectrum	5 user defined independent filter wheels in the range 350-750 nm	4 channels (450-495, 500-550, 565-590, 630-650)
Filters/detection channels	5 user defined independent filter wheels in the range 350-700 nm (default: FAM, Hex, ROX, Cy5, Cy 3)	4 channels (510-527, 565-590, 606-650, 670-750) (default: FAM, Cy3, TET, Alexa 532, Texas Red, Cy5, Alexa 647 and Eva Green)
Format	96-well plates, 0.2 ml tubes, 8 x 0;2ml strips	16 proprietary tubes
Time (40 cycles)	< 1.5 hours	20 - 40 mins
Reaction Volume	10-50 μl	25 or 100 μl
Fluorescence Chemistry	hydrolysis probes, fret probes, molecular beacons, Scorpions, Amplifluor, SYBR Green I	hydrolysis probes, molecular beacons, Eclipse probes, Amplifluor and Scorpion primers, SYBR Green I
Multiplexing Supported	5-plex	4-plex
Passive Reference	Optional ROX	Not required
Dimensions (H x W x D)	33 x 46 x 43 cm	30 x 30 x 25 cm
Weight	20 Kg	10 Kg
Other features	PCR mastermix kits	PCR mastermix kits and parameter specific kits
	System compatible with 21 CFR part 11 requirements	Random access (16 independently programmable reactions site)
	Software: gene expression analysis, allelic discrimination, melting curve analysis	Software: gene expression analysis, allelic discrimination, melting curve analysis and endpoint
	Integrated computer	Compact and portable for field use
		Modular system - flexibility to expand in multiples of 16 module units up to 96 reaction sites

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	Corbett	Eppendorf
Model	Rotor-Gene 6000	Mastercycler ep realplex⁴
Laser/Lamp	6 high power LEDs	96 LEDs
Detector	PMT	2-channel PMT
Thermocycling	Resistive heater with air cooling and centrifugation	Gradient peltier element block
Excitation spectrum	365, 470, 530, 585, 625, 680 nm,	470 nm
Filters/detection channels	460, 510, 555, 610, 660, 712 nm	520 / 550 / 580 / 605 nm
Format	36 0.2 ml or 72, 0.1 ml plastic tubes; 72 or 100 well Gene-Disc plastic rings	96-well plates, 0.2 ml tubes
Time (40 cycles)	40 mins	30 mins
Reaction Volume	10-100μl, 10-50μl, 15-50μl, 10-25μl	5-100μl
Fluorescence Chemistry	hydrolysis probes, molecular beacons, hybridisation probes, SYBR Green I	Open system all formats supported, except hybridisation probes
Multiplexing Supported	6-plex	4-plex
Passive Reference	Not required	Not required
Dimensions (H x W x D)	27.5 x 37 x 42	39.6 x 26 x 41 cm
Weight	14 Kg	24 Kg
Other features	Centrifugal rotary design	PCR mastermix kits
	Software: HRM (high-resolution melt), end-point, association / disassociation (i.e. melting / annealing) kinetics, relative expression and nucleic acid concentration measurement	Software: raw data, melting curve analysis, quantification/relative quantification, allele discrimination, endpoint, +/- assays.
		Gradient range of 24°C

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	BioRad	BioRad
Model	MiniOpticon	MyiQ
Laser/Lamp	48 LEDs	Tungsten-halogen lamp
Detector	2 photodiodes	12-bit CCD camera
Thermocycling	MJ Mini peltier element block	iCycler peltier element block
Excitation spectrum	470-500 nm	475-495 nm
Filters/detection channels	523-543 nm 540-700 nm	515-545 nm
Format	48 well PCR plate, 6 x 8 tube strips	96 well plates, 0.2ml tubes
Time (40 cycles)	< 40 mins	< 40 minutes
Reaction Volume	10-100 μl (20 μl recommended)	10 - 100 μl
Fluorescence Chemistry	hydrolysis probes, molecular beacons, SYBR Green I	hydrolysis probes, molecular beacons, SYBR Green I
Multiplexing Supported	2-plex	Single-plex
Passive Reference	Not required	Fluorescein
Dimensions (H x W x D)	33 x 18 x 32	39 x 29 x 58
Weight	6.8 Kg	17.6 Kg
Other features	PCR mastermix kits	PCR mastermix kits
	Software: allelic discrimination, gene expression analysis, melting curve analysis	Software: allelic discrimination, gene expression analysis, melting curve analysis, end point detection, absolute quantification
	Connect 4 instruments to 1 PC	Optional software for 21 CFR part 11 compliance
	Gradient range of 16°C	Gradient range of 25°C

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	BioRad	BioRad
Model	Opticon2	Chromo4
Laser/Lamp	96 LEDs	4 LEDs in photonics shuttle
Detector	Dual PMTs	4 photodiodes
Thermocycling	DNA Engine peltier element block	DNA Engine Dyad Disciple peltier element block
Excitation spectrum	470-505 nm	450-650 nm
Filters/detection channels	523-543, 540-700 nm	515-730 nm
Format	96-well plate, 8 x 0.2ml strip tubes	96-well PCR plate or 96 x 0.2 ml tubes
Time (40 cycles)	< 40 mins	< 40 mins
Reaction Volume	10-100 μl (20 μl recommended)	10-100 μl (20 μl recommended)
Fluorescence Chemistry	hydrolysis probes, molecular beacons, SYBR Green I	hydrolysis probes, molecular beacons, SYBR Green I
Multiplexing Supported	2-plex	4-plex
Passive Reference	Not required	Not required
Dimensions (H x W x D)	60 x 34 x 47 cm	37 x 24 x 35
Weight	29 Kg	12.6 Kg
Other features	PCR mastermix kits	PCR mastermix kits
	Software: allelic discrimination, gene expression analysis, melting curve analysis	Software: allelic discrimination, gene expression analysis, melting curve analysis
	Gradient range of 24°C	Customisable filter sets
		Modular design for maximal flexibility, 2 x 96 Dyad
		Gradient range of 24°C

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

Company	BioRad
Model	iQ5
Laser/Lamp	Tungsten halogen lamp
Detector	12-bit CCD camera
Thermocycling	iCycler peltier element block
Excitation spectrum	5 filter positions in range 475-645 nm
Filters/detection channels	5 filter positions available in range 515-700 nm, 2 provided
Format	96- or 384-well plate, 8 x 0.2ml strip tubes
Time (40 cycles)	< 40 minutes
Reaction Volume	15-100 μl
Fluorescence Chemistry	hydrolysis probes, molecular beacons, hybridisation probes, SYBR Green I
Multiplexing Supported	5-plex
Passive Reference	Fluorescein
Dimensions (H x W x D)	36 x 33 x 62 cm
Weight	17.6 Kg
Other features	PCR mastermix kits
	Software: allelic discrimination, gene expression analysis, melting curve analysis, end point detection, absolute quantification
	Gradient range of 25°C
	Optional software for 21 CFR part 11 compliance

Abbreviations: LED, light emitting diode; CCD, charge-coupled device camera; PMT, photomultiplier tube; UPL, universal probe library

from Julie Logan, Kirstin Edwards and Nick Saunders in Real-Time PCR: Current Technology and Applications

	Real-Time PCR: Advanced Technologies and Applications Edited by: Nick A. Saunders and Martin A. Lee Provides both the novice and experienced user with an invaluable reference to a wide-range of real-time PCR technologies and applications and supplies detailed technical insights into the underlying principles, methods and practice of real-time PCR. read more ...
	Real-Time PCR in Food Science: Current Technology and Applications Edited by: David Rodríguez-Lázaro An indispensable manual on real-time PCR for scientists in the food industry and for anyone involved in the detection of foodborne pathogens. read more ...
	Quantitative Real-time PCR in Applied Microbiology Edited by: Martin Filion Aimed specifically at microbiologists, this volume describes and explains the most important aspects of current real-time quantitative PCR (qPCR) strategies, instrumentation and software. "useful book ... filled with valuable information" (Doodys) read more ...
	PCR Troubleshooting and Optimization: The Essential Guide Edited by: Suzanne Kennedy and Nick Oswald Control, optimize and troubleshoot PCR, reverse transcriptase PCR, real-time PCR and quantitative PCR. An essential book. "an essential book ... a valuable tool to all those interested in PCR" (Doodys); "an essential guide" Aus. J. Med. Sci. read more ...
	Lab-on-a-Chip Technology (Vol. 2): Biomolecular Separation and Analysis Edited by: Keith E. Herold and Avraham Rasooly Applications in the biomedical and life sciences: biomolecule separation, electrophoresis, chromatography, protein and cell separation, genetic and transcriptome analysis, PCR, cell viability analysis and microorganism capturing. "a comprehensive and felicitous compendium" (Drug Research) read more ...