Introduction to Piko^® thermal cycling technology

Piko demonstration movie
Uniformity of the Piko Thermal Cycler
Fast block temperature settling
High yields and uniform results
High speed and yield
More product in less time
Low volume PCR
Peltier technology

Finnzymes’ high-quality Piko® Thermal Cyclers represent the next generation of PCR instrumentation. To help researchers achieve significant improvements in DNA amplification such as protocol speed, consistency of results and overall running cost, Finnzymes has developed a novel technology termed “Piko.” The Piko Thermal Cycler is based around the Piko PCR Plate which is ¼ the size of conventional microplates yet maintains industry-standard sample capacity and well-to-well spacing routinely used in research and diagnostic labs. This smaller plate format has enabled the design of a PCR instrument with remarkable improvements: the Finnzymes’ Piko Thermal Cycler is extremely fast, very tiny and highly economical to use.

Piko^® demonstration movie

Click on a link below to download a quicktime demonstration movie (Requires Apple QuickTime 7).
Piko demonstration movie medium resolution (5.7 MB)

 

Excellent temperature uniformity across the block

Piko Thermal Cycler at 60°C		Competitor's thermal cycler at 60°C
Piko Thermal Cycler at 95°C		Competitor's thermal cycler at 95°C
Figure 1. Temperature uniformity Block temperature uniformity of the 96-well Piko Thermal Cycler and another manufacturer’s fast 96-well thermal cycler were measured at two commonly used temperatures for PCR. Remarkably, even at 95°C the Piko Thermal Cycler shows a maximum temperature difference between any two wells to be less than 0.4°C, as compared with a top competitor’s systems of greater than 0.8°C. Steady-state temperature measurements were performed with a 16-probe MTAS™ temperature verification system.

 

Piko^® Thermal Cyclers' block temperature settling time is only 1 second

The Piko Thermal Cyclers’ best-in-class thermal performance is characterized by near-perfect uniformity and 1-second settling times (the time it takes for the temperature of all samples in the block to reach equilibrium).

Figure 2. Block temperature settling time The temperature of each well was individually measured during block temperature ramp to 98°C (blue lines). The steep slope of the non-uniformity trace (solid red line) indicates extremely fast settling time across the block. Measurements were performed with a 16-probe MTAS™ temperature verification system from Cyclertest.

 

High yields and uniform results in 10 minutes.

Figure 3. High yields with a fast protocol
A 400 bp amplicon from lambda DNA was amplified with an ultra-fast PCR protocol on a 24-well Piko Thermal Cycler using ultra-thin wall (UTW®) reaction vessels and Phusion® Flash High-Fidelity PCR Master Mix. 5 μl aliquots of each 15 μl reaction were run on an agarose gel. The following PCR protocol was used: 25 cycles of [98°C for 0 sec, 72°C for 0 sec], 72°C for 10 sec. Total cycling time was 10 min.

 

High speed and yield with the Piko^® Thermal Cycler

Total protocol time	15 µl < 14 minutes 50 µl < 15 minutes
	98°C 0 s	} 35x
Protocol	72°C 0 s
	72°C 30 s

Figure 4. High speed PCR with human genomic DNA in under 15 minutes
A 581 bp human genomic sequence (glutathione peroxidase-3) was amplified in replicate using UTW® vessels and Phusion® Flash DNA polymerase with a 24-well Piko® thermal cycler.  Note that across the range of volumes (15 – 50 µl) the uniformity and yields are high with a minimal amount of variation from sample to sample. This is due to the Piko’s fast settling time and ultra-thin wall vessels that allow the sample to come to steady-state temperature quickly.  Phusion provides higher yields by utilizing both its inherent high processivity and its maintenance of high activity throughout the protocol due to its resistance of denaturation at high temperatures.  Total time for this protocol is under 14 minutes for the 15 µl sample and less than 15 minutes for the 50 µl sample.

 

More product in less time as compared with conventional PCR

Figure 5. More product in less time
A 2kb fragment from the Cathepsin K gene and a 245bp fragment from the ApoB gene was amplified in triplicate using 3-step PCR protocols optimized for either Phusion® Flash DNA polymerase with a Piko® thermal cycler or Taq DNA polymerase with a Tetrad™ thermal cycler. The Finnzymes High Performance PCR system offers a 69% time savings and much higher yields.

  

Low volume PCR with the 96-well Piko^® Thermal Cycler

Figure 6. Low volume PCR
A 400bp lambda DNA and a 695bp human β-Glucuronidase DNA fragment were amplified in 5µl reaction volumes using UTW® vessels and Phusion® DNA polymerase with a 96-well Piko® thermal cycler.  The thermal cycling parameters were 98° for 0 seconds and 72° for 5 seconds (400bp) or 15 seconds (695bp), for 35 cycles, followed by a 30-second final extension. 4µl of product were loaded onto the gel according to the grid below. Note that the uniformity and yields are high with a minimal amount of variation from sample to sample. The total times for each protocol were 14 minutes 38 seconds and 20 minutes 37 seconds, respectively.

96-well Piko® PCR Plate representation showing which wells were loaded onto the above gels.

 

Peltier technology

Download the Peltier technology movie (1.7 MB)

At the heart of the Piko cycler are the Peltier elements that provide precise control of the heating and cooling where the reaction vessels are placed. The Peltier elements are comprised of many pairs of p and n semi-conductor dice. To initiate the movement of heat, electrical current is applied and this alone results in the removal of heat from one side of the element and the depositing of the heat to the opposite side of the element – in effect a heat pump. Even though electrons flow through each pair of dice in opposite directions, they synchronously pump heat in the same uniform direction. By reversing the polarity of the voltage through these junctions, the electron flow is reversed. This changes the direction of heat flow resulting in either a heating or cooling effect to the sample.