qPCR Machines and Real-time PCR Platforms
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.
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
Further reading
- Real-Time PCR: Advanced Technologies and Applications
- Real-Time PCR in Food Science: Current Technology and Applications
- Quantitative Real-time PCR in Applied Microbiology
