Speed of PCR Research Publications @ dna-utah.org

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).