GspSSD2.0 LF DNA Polymerase
Key features:
Cat. No: | Description | |
---|---|---|
GSPSSD2-001 | 1600u | GspSSD2.0 DNA polymerase @ 8u/µl |
GSPSSD2-002 | 8000u | GspSSD2.0 DNA polymerase @ 8u/µl |
GSPSSD2-002HC | 8000u | GspSSD2.0 DNA polymerase @ 100u/µl |
GSPSSD2-003HC | 40000u | GspSSD2.0 DNA polymerase @ 100u/µl |
iBuffer5-001 | 2 x1.5ml iBuffer 5 | |
iBuffer5-002 | 15ml iBuffer 5 |
GspSSD2.0 vs competitor ‘N’ (LAMP Pol 3.0) and competitor ‘L’ (thermostable LAMP Pol)
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.
GspSSD2.0 vs competitor ‘N’ (LAMP Pol 3.0) and competitor ‘L’ (thermostable LAMP Pol)
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 MS2 primer 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.