SOLIS BIODYNE OÜ A: Teaduspargi 9, 50411 Tartu, Estonia E: [email protected] T: +372 7409 960 SolisFAST® Lyo-ready qPCR Kit with UNG solisbiodyne.com SolisFAST® Lyo-Ready qPCR Kit with UNG represents a glycerol-free qPCR solution with optimized excipients that is suitable for reliable lyophilization. The Kit is designed for fast and sensitive amplification and quantification of DNA targets using probe-based assays. The formulation is tailored for effective freeze-drying to produce stabilized cakes or beads that easily dissolve upon reconstitution with sample material. With a high collapse temperature (Tc) of -30.8 ˚C, the kit allows for the adoption of a rapid and energy-efficient lyophilization protocol. With a glass transition temperature (Tg) of 68.3˚C, the lyophilizates have a high resistance against long term exposure to higher temperatures. Lyophilizing together with primers and probes provides maximum convenience for further reaction set up. Inhibitor tolerance and fast extension rates of the SolisFAST® DNA Polymerase enable quick and robust DNA detection even from complex biological sample types. The Kit comes in a flexible 2 tube format: a glycerol-free qPCR mix (5x) and a lyophilization excipient mix (4x). SolisFAST® Lyo-Compatible qPCR Mix with UNG offers you: SolisFAST® Lyo Excipient Mix gives you: • Flexibility with lyophilization – the qPCR mix formulation is glycerol free, making it suitable for freeze-drying. Either use it in combination with Solis BioDyne’s proprietary SolisFAST® Lyo Excipient Mix (supplied with the kit) or opt for your own additives • Sensitive multiplex detection – detect low copy numbers with up to 5-plexing • Short run times - fast extension rates of the SolisFAST® DNA Polymerase enable you to save time by using quick cycling protocols • Peace of mind with UNG – Salini UNG® Uracil-N-Glycosylase will eliminate carryover contamination and prevent false positive results • Efficient lyophilization into cakes and beads • Strong cryoprotection during freezedrying • Stabilized lyophilizates with high resistance against long term exposure to higher temperatures • Rapid and seamless reconstitution • Preserved qPCR performance after lyophilization Figure 1. ALB target from human gDNA was amplified over six 10-fold dilutions (500 ng to 5 pg; E=104%), showing sensitive detection over a wide dynamic range. Reactions were run on Bio-Rad CFX96 platform. Prevent false positive results with Salini UNG® Uracil-N-Glycosylase Amplification 6000 5000 4000 RFU Wide dynamic range Liquid form Lyophilized with primers and probes Lyophilized form Figure 2. Amplification plot showcasing results obatained with SolisFAST® Lyo-Ready qPCR Kit with UNG in liquid form (red), lyophilized form (blue), lyophilized with primers and probes (green) and a formulation without UNG (orange). All reactions were spiked with an equal concentration of dU-containing amplicons, mimicking carryover contamination. UNG maintained full functionality in all conditions, degrading all the amplicons, while control reactions without

SolisFAST® Lyo-Ready qPCR Kit with UNG shows consistent performance before and after lyophilization, as well as after lyophilization with primers and probes. Cy5 Figure 3. 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...