Real-time PCR

Reducing cost through assay miniaturization and non-contact liquid-handling.

Introduction

Real-Time PCR or quantitative PCR (qPCR) is a powerful method for detecting and quantifying specific RNA or DNA sequences. Preparation of multiwell plates for qPCR analysis can be very time consuming and prone to mistakes when liquids are distributed manually with a single pipette. Multichannel pipettes and modern pipetting robots may decrease the preparation time but are very limited in terms of combining multiple source liquids in a certain pattern as it is required for the standard qPCR assay. Furthermore, conventional liquid handling technologies are only able to dispense reliable volumes starting from about 1 µL or require large volumes of the source liquid as a reservoir for dispensing, which has to be discarded and can’t be reused after plate preparation.

How the I.DOT facilitates real-time PCR

Dispendix’s portfolio features the Immediate Drop-on-demand Technology (I.DOT), a patented approach for optimizing noncontact liquid-handling tasks. The I.DOT supports multiple liquid classes and can dispense a variety of solutions, including PCR buffer, DMSO and glycerol up to 50%, making it compatible with both commercial and in-house generated reagents. Accurate dispensing is essential for producing high-quality data that shows minimal technical variation between samples. The I.DOT’s built-in droplet verification detects and counts every droplet released during a single dispensing run. The I.DOT is the only liquid dispenser that offers built-in droplet quality control.

REDUCING COST PER DATA POINT

Because of expensive reagents, the cost for qPCR assays fluctuates with the number of samples and the working volume. Assay miniaturization with our I.DOT low-volume liquid-handling technology significantly reduces cost for reagents, by allowing precise reagent dosing in the nL volume range. qPCR reactions can be easily scaled down to sub-μL volumes in 384- or 1536-well plates. In addition, the I.DOT does not require pipette tips.

REDUCING TIMELINES AND INCREASING REPRODUCIBILITY

With the I.DOT, the preparation of a 384-well plate for qPCR can be performed in under 5 minutes. Multiple liquid combinations and volumes can be assembled using eight dispensing channels at the same time. In addition to the potential for countless combinations and fast performance, the noncontact dispensing technology can be easily integrated into automated workflows that use plate-handling robots. For even higher throughput, 1536- and 3456-well plates can be used.

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