KleverLab introduces lyo-ready and lyophilized Fast Probe qPCR mastermixes

We are expanded FastProbe products line with new lyo-Ready and lyophilized qPCR mastermixes. The new portfolio includes lyophilized Fast Probe Lyo qPCR Mastermix and Fast Probe U Lyo qPCR Mastermix which can be stored at room temperature and 5X Fast Probe Lyo-Ready qPCR Mastermix with 5X Fast Probe U Lyo-Ready qPCR Mastermix for in-house production of fully lyophilized PCR mixes and kits.

Comparison of PCR results obtained with 5X Fast Probe Lyo-Ready qPCR Mastermix (green), mastermix after lyophilization and subsequent reconstitution (red) and mastermix after lyophilization, subsequent reconstitution and 10 freeze-thaws cycles of reconstituted mixture (blue).

Figure 1. Tetra-plex qPCR reactions were performed with the 5X FastProbe Lyo-Ready qPCR Mastermix and primers for detection GMO in soybeans (amplification plot from FAM channel is presented). The Mastermix was tested before lyophilization in liquid form (green), after lyophilization to cake (red) and after lyophilization to cake with 10 freeze-thaws cycles after reconstitution (blue). All channels and conditions produced consistent results: Cq values of respective DNA dilutions stayed within ±1,0 range, fluorescence levels did not fluctuate by more than 15% and efficiencies remained between 90-110%. The reactions were performed with plasmid DNA (50 and 10 copies in reaction) and run on Bio-Rad CFX96 platform using the following protocol: 2 min 95°C, 45 cycles of 5 sec 95°C, 15 sec 60°C, 15 sec 72°C.

Results of PCR obtained with Fast Probe Lyo qPCR Mastermix lyophilized with primers for AFSV detection and stored 39 months at room temperature (green) in comparison with freshly prepared Fast Probe mastermix (red).

Figure 2. 39 months storage of lyophilized FastProbe Lyo qPCR Mastermix with ASFV primers. Dual-plex qPCR reactions were performed with freshly made and lyophilized mastermixes (amplification plot from FAM channel (ASFV) is presented). The freshly made mastermix was tested in liquid form (green), stored lyophilized to beads with primers mix was reconstituted after 39 months at room temperature (red). All channels and conditions produced consistent results: Cq values of respective DNA dilutions stayed within ±1,0 range, fluorescence levels did not fluctuate by more than 15% and efficiencies 100,4% for liquid and 103,7% for lyophilized form. The reactions were performed with viral ASFV DNA (1000 and 5 copies in reaction) and run on Bio-Rad CFX96 platform using the following protocol: 2 min 95°C, 45 cycles of 5 sec 95°C, 15 sec 60°C, 15 sec 72°C.

You can order these PCR-products on our website now.