Introducing SolisFAST® Lyo-Ready qPCR Kit with UNG

There is a rising demand for lyophilized assays, and Solis BioDyne is delighted to address this by supporting their customers in achieving maximum user convenience with our newest product.

• The product features a glycerol-free qPCR mix, designed for probe-based qPCR, coupled with a carefully optimized lyophilization excipient mix. The two-tube format gives extra flexibility and allows you to work with your own choice of excipients should that be necessary for your lyophilization procedures.
• The solution is suitable for lyophilization to produce highly stabilized lyo-beads and lyo-cakes, with or without primers/probes, easily reconstituted with sample material.
• Comprising the SolisFAST® DNA Polymerase, the kit enables fast and sensitive 5-plex amplification in approximately 30-45 minutes.
• The incorporation of UNG ensures the elimination of false positive results caused by carryover contamination.

SolisFAST® Lyo-Ready qPCR Kit with UNG can easily compete with other similar products on the market. With a high collapse temperature (Tc) of -30.8 °C the kit allows for the adoption of a rapid and energy-efficient lyophilization protocol. Lyophilizing together with primers and probes provides maximum convenience for further reaction setup. Inhibitor tolerance and fast extension rates of the SolisFAST® DNA Polymerase enable quick and robust DNA detection even from complex biological sample types. With a high glass transition temperature (Tg) of 68.3 °C, the lyophilized product also has a high resistance against long-term exposure to higher temperatures. Stability tests show that the lyophilized products are stable for at least 6 months at plus 25 °C and have a very low residual moisture of 1.4 %.

Of course, one of the most important aspects of a lyophilized product is the ability to remain as functional after lyophilization as it was before. Below you can see a set of tests that were done with the product to demonstrate just that. The results clearly show that the kit ensures consistent performance with persistent Cq values before and after lyophilization, as well as after lyophilization with primers and probes.

Figure 1. Five-plex RT-qPCR reactions were performed with the SolisFAST® Lyo-Ready qPCR Kit with UNG (amplification plots from VIC and Cy5 channel are presented). The Kit was tested before lyophilization in liquid form (red), after lyophilization to cakes (blue), and after lyophilization to cakes with primers and probes (green). All channels and conditions produced consistent results: Cq values of respective DNA dilutions stayed within ± 0,5 range, fluorescence levels did not fluctuate by more than 15% and efficiencies remained between 90-110%. The reactions were performed with human gDNA (50 ng to 50 pg) and run on Bio-Rad CFX96 platform using the following protocol: 3 min 95 °C, 45 cycles of 3 sec 95 °C, 10 sec 60 °C.

Given the inclusion of the SolisFAST® DNA Polymerase in this product, which has very fast extension rates, tests were also conducted to evaluate its performance with both standard and fast cycling protocols in both liquid and lyophilized forms. The results show that the Cq values obtained with all the tested conditions are very persistent. Therefore, it can be confidently said that SolisFAST® Lyo-Ready qPCR Kit with UNG can produce consistent values before and after lyophilization with both standard and fast cycling protocols.

Consistency in Cq values with liquid and lyophilized reagents under standard and fast cycling

Figure 2. Five-plex qPCR reactions were performed with liquid and lyophilized SolisFAST® Lyo-Ready qPCR Kit with UNG using the standard and fast cycling protocol with four 10-fold DNA dilutions (from 5 to 0,005 ng/µl). Results from FAM channel are presented. The Cq values of respective DNA dilutions are consistent across all tested conditions. Standard cycling protocol: 95°C 3 min; 45 cycles of 95°C 5 sec, 60°C 20 sec. Fast cycling protocol: 95°C 3 min; 45 cycles of 95°C 1 sec, 60°C 5 sec.