Pipette White Paper: Thermal Cycling Module

Opentrons Thermal Cycling Module GEN2 provides accurate and uniform amplification temperatures for your experiments while maintaining low evaporation rates.

Product Description

The Opentrons thermal cycler module is a fully automated thermal cycler placed on the deck, providing a 96-well plate-style fully automated PCR function. The thermal cycler module is compatible with all Opentrons instruments and software, so fully automated PCR pre-treatment and PCR processes can be implemented on Opentrons instruments.

Opentrons The well plate of the thermal cycle module is equipped with an automatic heating cover and a replaceable sealing element to ensure effective heating of the sample and evaporation of the sample< /strong> Reduce the quantity to a minimum. After the thermal cycle is completed, the well plate blockis raised through the well plate lifting device, and then the PCR well plate Move out. The exhaust vent removes hot air from the thermal cycler and directs it away from the machine deck.

Performance Data

Opentrons conducted a series of rigorous performance tests on the thermal cycle module GEN2 to ensure that the quality of the instrument complies with industry standard performance specifications

Table 1: Accuracy and consistency of temperature at steady state

Temperature rise and fall rate (temperature change rate)

The average maximum temperature rise and fall rates were measured for 8 different instruments during heating (to 99°C) and cooling (to 4°C). The holding time of each fixed point is 5 minutes. The maximum average temperature change rate of the thermal cycler is 4.45℃/s

Performance Data

The instrument's amplification consistency was tested by using reusable sealing elements, multiple reaction systems, and 96-well PCR plates. Use Lambda primers to amplify Lambda DNA. The thermal cycle parameters are as follows: incubate at 94°C for 10 minutes, 25 cycles, each cycle amplifies at 94°C for 15 seconds and 70°C for 1 minute, and incubates at 72°C for 15 seconds. Extend for 7 minutes. The amount of amplification in each well was measured by capturing raw fluorescence values ​​using a read trigger. The coefficient of variation (CV%) is calculated and shown in Table 2. Data shows that the seal can be reused up to 25 times without loss of performance. CV values ​​remained low at both 10 μL and 100 μL reaction volumes. In addition, using the OT-2 96-well PCR plate, the thermal cycle module has good amplification repeatability in the large range of 10uL to 100uL.

Low evaporation rate

The evaporation rates of 50 different thermal cycler modules were evaluated with sample volumes of 10 μL, 20 μL, 50 μL, and 100 μL in a PCR 96-well plate. The volumetric weight of the sample was measured before and after the 3 steps of PCR operation; including 35 cycles, the set temperature and holding time of each cycle were: 94°C (10 s), 70°C (30s) and 72°C (30s) .

The average evaporation rate of all tested sample volumes was <6%, and the average evaporation rate of the largest sample volume (100 μL) was <1%.

The data shows that the Opentrons thermal cycling module GEN2 has consistent and reliable amplification capabilities with minimal sample volume loss as low as only 10 μL. The model demonstrates rapid temperature rise and fall rates and the ability to maintain stable temperature at a variety of different set points. Amplification in the 96-well plate was demonstrated to be consistent and uniform when disposable seals were reused, multiple reaction volumes were used, and regardless of whether samples were prepared in-situ. Opentrons Thermal Cycler GEN2's 9-6-well PCR has been proven through various tests to provide reproducible and accurate amplification results to ensure the consistency of daily laboratory experiments.

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