This mastermix incorporates the patented highly thermostable Tin(exo-) LF DNA Polymerase, renowned for its extreme thermostability and robust strand-displacement activity, enabling applications previously unattainable in a single, closed-tube reaction with alternative isothermal enzymes.
Featuring extreme thermostability* below 98ºC, robust strand displacement activity, and optimized performance at 65ºC, this mastermix stands out with 5’-3’ exonuclease minus and 3’-5’ exonuclease minus properties, ensuring high fidelity throughout the amplification process.
*no loss of activity is observed in a LAMP reaction following 5 minutes @ 95ºC step.
Employing a template denaturation step at 95ºC prior to amplification has been demonstrated to enhance target sensitivity[1], while also enhancing primer specificity, thereby reducing false-priming occurrences common at lower temperatures. Furthermore, the process offers straightforward template extraction through cell lysis, enabling sample heat lysis, amplification, and detection within a single tube, streamlining the workflow.
[1] Mori Y, Notomi T. 2009. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases. J Infect Chemotherapy. 15(2):62-9.
| Cat. No: | Primary Application |
| ISO-001Tin | Tin DNA polymerase Isothermal Master Mix - 300 reactions |
| ISO-DR001Tin | Tin DNA polymerase Isothermal Master Mix - 300 reactions - Dried |
| ISO-002Tin | Tin DNA polymerase Isothermal Master Mix - 1500 reactions - 5 x 300rxn kits |
Effect of heat-denaturation on ISO-001Tin LAMP (Human gDNA template): Time-to-threshold comparison:
25μl HD-LAMP reactions: 8U DNA Pol, 1X manufacturers supplied reaction buffer (‘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. ISO-001Tin and ‘L’ reactions heated at varying temperatures for 5 minutes prior to amplification as per manufacturer’s instructions at 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.
ISO-001Tin vs competitor ‘N’ (LAMP Pol 3.0) and competitor ‘L’ (thermostable LAMP Pol)
