OptiGene/GspSSD2.0 LF DNA Polymerase/GspSSD2.0 DNA polymerase @ 8u/µl/1600u/GSPSSD2-001

LAMP reactions (25µl): 8U supplied DNA Pol, 1X manufacturers supplied reaction buffer (‘GspSSD2.0’ – iBuffer5, ‘N’ – IsoPol II, ‘L’ – BufferC), dNTPs/MgSO4 and Betaine – as per manufacturers recommended concentrations, 100-10,000 copies Human gDNA (Sigma), 1X Human VKORC1 primer mix – designed using LAMPDesigner™ (0.8µM FIP 5’-CTGTTGCACCTACCCTTCTGGAGCAGGAGAG GGAAATATCA-3’, 0.8µM BIP 5’-GAAGTCAAGCAAGAGAAGACCT GAAAACTCCTGACCTCAAGTGA-3’, 0.2µM F3 5’-ACCATTATTCTGTCTACCACAC-3’, 0.2µM B3 5’-GACAGGGTTTCACCATGTT-3’, 0.4µM LoopF 5’-TCTCTTCCTCTGGCGTCT-3’, 0.4µM LoopB 5’-CTCACGCCTATAATCCTAGCATT-3’), 3.75ng fluorescent intercalating dye (V13-01184 Dyomics GmbH).‘GspSSD2.0’, ‘N’ and ‘L’ reactions were run as per manufacturer’s instructions at 65ºC, 65ºC and 70ºC respectively on a Genie®II.A: Real-time amplification plots. B: Time-to-threshold values (the maximal rate of change in fluorescence over time) reported in graphical format for simple comparison.

LAMP reactions (25µl): 8U supplied DNA Pol, 1X manufacturers supplied reaction buffer (‘GspSSD2.0’ – iBuffer5, ‘N’ – IsoPol II, ‘L’ – BufferC), dNTPs/MgSO4 and Betaine – as per manufacturers recommended concentrations, 0.1-10ng MS2 RNA (Roche), 1X MS2primer mix designed using LAMPDesigner™(0.8µM FIP 5’-TCGCAAGCGAACCATCTACGTCCGTTGGTATAGACCTGAA-3’, 0.8µM BIP 5’-CGATAGACTTATCGTCTGCATCCGGAGATATGAATATAGCTCTGGTGG-3’, 0.2µM F3 5’-TGTAAGGAGCCTGATATGAATATG-3’, 0.2µM B3- TAGTGTGAGCGGATACGAT-3’, 0.4µM LoopF 5’-GCCAGACGCTGGTTGAT-3’, 0.4µM LoopB 5’-GATCGCCTGGTGTGGAG-3’), 3.75ng fluorescent intercalating dye (V13-01184 Dyomics GmbH).‘GspSSD2.0’, ‘N’ and ‘L’ reactions were run as per manufacturer’s instructions at 65ºC, 65ºC and 70ºC respectively on a Genie®II +/- 0.5U AMV-RT (OptiGene).A: Real-time amplification plots. B: Time-to-threshold values (the maximal rate of change in fluorescence over time) reported in graphical format for simple comparison.