Characteristics of studies with reported PCR cycle times of 10 seconds or faster

Reproduced & Updated from Wittwer CT et al. 2025, PMID: 40260633

Fastest PCR Reported (s/cycle)
System Design
Heating Method
Cooling Method
Template
[Template] (copies/µL)
Product Length (bp)
Polymerase
[Polymerase] (nM) *
[Primer] (nM)
Negative Control Used?
Assessment of Amplification
Quality of Amplification
Group
0.42
Capillary tube moved between 2 water baths via motor
Water
Water
Human
3000
60
Klentaq
8000
20000
Yes
Real-time monitoring
Robust
2015 Farrar and Wittwer
1.05
Flow-through (oscillating)
Thin-film platinum resistor within microchannel
Passive cooling (heat sink)
Synthetic
5000
70
Klentaq
3500
5000
Yes (not shown)
Melting curve analysis
Robust
2019 Myrick et al.
1.06
Flow-through
Copper block
Copper block
Synthetic
2100
69
Klentaq
2000
5000
Yes
Melting curve analysis
Moderate
2017 Trauba and Wittwer
1.8
Radiative using Au nanoparticles
Radiative (IR laser)
Chilled airflow
Plasmid
NA
79?
KAPA2G
4
500
Yes (low signal)
Endpoint fluorescence
Weak
2017 Roche et al.
2
Flow-through
Copper block
Copper block
Human
1500
102
Klentaq
64
2000
Yes
Real-time monitoring & melting
Moderate
2018 Jafek et al.
2.6
Pressurized gas and capillaries
Forced air convection
Forced air convection
Bacteria
360
85
Taq
100
500
No
Electrophoresis gel
Weak
2004 Whitney
2.7
Capilary tubes in gallium eutectic
Peltier
Peltier
Bacteria
NA
85
KOD
NA
500
No
Electrophoresis gel
Weak
2010 Maltezos et al.
3
Flow-through
Aluminium
Passive cooling (aluminium)
Bacteria
10000
134
Ex Taq
13
200
Yes (low signal)
Real-time monitoring
Weak
2011 Fuchiwaki et al.
3
Flexible capillary moved between 2 water baths via motor
Water
Water
Soybean
0.03 - 300
165
SpeedSTAR
54
400
Yes
Electrophoresis gel & imaging
Moderate
2018 Wang et al.
3.7
Microfluidic chip
Peltier
Peltier with
Human cDNA
3800
89
Taq (HotStart)
50
NA (no primer info)
Yes (not shown)
Electrophoresis gel
Weak
2019 Lee et al.
4
Flow-through
Polyimide film heaters
Passive cooling
Leukemia cell line..
670
151
KlenTaq
6000
3000
No
Electrophoresis gel
Robust
2022 Lin et al.
4
Droplet PCR (in oli)
Radiative (IR laser)
Passive cooling
Bacteria
33 - 100
72
KOD
NA
NA
No
Real-time monitoring
Moderate
2008 Terazono et al.
4.3
Flow-through
Aluminium
Aluminium
Bacteria
125
Multiplex 197 & 311
SpeedSTAR
54
400
Yes (not shown)
Electrophoresis
Weak
2019 Li et al.
4.6
Convective heat transfer through porous copper media with resistive heating
Water
Water
Bacteria
190000
160
KAPA2G
9
400
Yes (not shown)
Electrophoresis gel
Moderate
2011 Wheeler et al.
4.8
Silicon chip on Peltier
Peltier
Peltier
Bacteria
4.2 - 4200
78
KAPA2G
370
20000
Yes
Real-time monitoring & melting
Robust
2019 Cai et al.
5
Capillary tube moved between 2 water baths
Water
Water
Maize
NA
195
Taq
300
2000
Yes
Electrophoresis gel
Moderate
2019 Gao et al.
5
Digital droplet PCR between two water baths
Water
Water
Bacteria
0.001 – 1000
192
SpeedSTAR
86
400
Yes
Digital endpoint fluorescence
Moderate
2023 Wang et al.
5.2
Flow-through ,(polycarbonate)
Resistance film heaters
Passive cooling
Lambda phage
200000000
500
Taq
14
200
No
Electrophoresis gel
Weak
2004 Hashimoto et al.
5.3
Water pumped against aluminum plate with 1µL droplets
Aluminium
Aluminium
Bacteria
2200000
72
KOD
24
250
Yes
Real-time monitoring
Robust
2010 Terazono et al.
5.6
Droplet shuttled between zones using electrowetting
ITO resistive heating
Passive cooling
Plasmid
170000
63
KAPA2G
22
500
Yes
Real-time monitoring
Moderate
2023 Wan et al.
5.7
Flow-through
Aluminium
Aluminium
Bacteria
NA
Multiplex 197 & 316
SpeedSTAR
1
200
Yes (not shown)
Electrophoresis
Moderate
2021 Li et al.
6.3
Flow-through
Aluminium
Aluminium
Plasmid
400
NA
Ex Taq
13
400
Yes (low signal)
Endpoint fluorescence
Moderate
2011 Fuchiwaki et al.
6.5
Photonic heating on a chip with TiN nanorings
Radiative (IR LED)
Passive cooling
Lambda phage
NA
100
Taq
NA
400
No
Electrophoresis gel
Weak
2024 Kim et al.
6.6
Flow-through (silica chip)
Copper block
Copper block
PCR product
10000000
176
Taq
50
1000
Yes
Electrophoresis gel
Weak
1998 Kopp et al.
6.6
Gold nanoislands on glass nanopillar arrays
Radiative (White LED)
Passive cooling
Lambda phage
9000
98
Taq
5
9000
Yes
Real-time monitoring
Robust
2021 Kang et al.
6.7
Primer-immobilized hydrogels containing graphene oxide
Radiative (NIR LED)
Passive cooling
Synthetic
10000000
59
Taq
NA
NA
Yes
Real-time monitoring
Robust
2022 Kim et al.
7
Gold nanoislands on glass nanopillar arrays
Radiative (White LED)
Passive cooling
Lambda phage
1800000
98
Taq
5
9000
Yes
Electrophoresis gel
Robust
2020 Lee et al.
7.5
Silicon chip
Peltier
Peltier
Pseudo virus
NA
116
Kapa2G
40
360
Yes
Real-time monitoring
Moderate
2022 Zhang et al.
8.5
Micromachined cantilever terminated in a disc
Thin silicon film heater
Passive cooling
Mouse cDNA
NA
82
AmpliTaq
NA
900
No
Real-time monitoring
Robust
2006 Neuzil et al.
9
Flow-through (circular chip)
Copper block
Passive cooling
Lambda phage
18000000
500
Taq
5
1000
Yes
Electrophoresis gel
Moderate
2007 Sun et al.
9
Droplets in oil
Radiative (IR laser)
Passive cooling
Human 18S rDNA
1000000
187
AmpliTaq
NA
900
No
Real-time monitoring
Robust
2009 Kim et al.
9.8
Plasmonic PCR with Au film
Radiative (Blue LED)
Passive cooling
Human cDNA
2000
116
KAPA2G
1100
1000
No
Electrophoresis gel
Weak
2015 Son et al.
10
Plasmonic PCR with Au film
Radiative (Blue LED)
Forced air convection
Lambda phage
1800000
98
Taq
5
900
No
Electrophoresis gel
Weak
2015 Son et al.
10
Capillary tube moved between 2 water baths
Water
Water
Bacteria
NA
Multiplex 123 & 284
ExTaq
380
2000
Yes
Electrophoresis gel
Robust
2018 Tian et al.
20
Air-cycling system using thin glass capillary tubes
Forced air convection
Forced air convection
Human
1500
536
Taq
16
500
Yes (not shown)
Electrophoresis gel
Robust
1990 Wittwer et al.


Notes: Ordered according to the fastest successful cycle time reported in the paper, whether through direct citation, or dividing the reported total time of amplification by the number of cycles. Additional time before or after amplification (such as incubation steps) were not included. The 1990 report of 20 second cycles is listed at the end. "Negative Control Used?" asks whether an appropriate no-template control (NTC) was used with the fastest protocol. “Yes” means that it was, and the target signal was not observed. “No” means that an NTC was not mentioned, and the reaction specificity is uncertain. “Yes (low signal)” means that fluorescence was detected in the NTC, and amplification was considered successful for the target if the fluorescence was above a threshold. "Flow Through" is defined as a microfluidic system that moves DNA solution through different zones of temperature, such as by serpentine channels

Abbreviations: Au (gold); Cq (quantification cycle; replaces Ct or equivalent); IR (infrared); ITO (indium tin oxide); LED (light-emitting diode); NA (information not available); NIR (near infrared); TiN (titanium nitride)

*Polymerases concentrations were either measured by absorbance at 280 nm (if pure enzyme was available) or converted from activity units according to Table 1 of Montgomery JL, Rejali N, Wittwer CT. Stopped-flow DNA polymerase assay by continuous monitoring of dNTP incorporation by fluorescence. Anal Biochem. 2013;441:133-139. (PMID: 23872003, DOI: 10.1016/j.ab.2013.07.008). For heat stable enzymes like those used in PCR, concentrations are easier to compare than unit definitions that are often variable. Polymerase enzymes (vendors): AmpliTaq (Thermo Fisher); ExTaq (Takara bio); KAPA2G (KapaBiosystems); KlenTaq (DNA Polymerase Technology); KOD (Toyobo); SpeedSTAR (TakaraBio); Taq (multiple sources).