OptiGene’s patented thermostable DNA polymerase for amplification reactions requiring an initial heat-denaturing step. Extreme thermostability and strong strand-displacement activity enable applications unachievable in a single, closed-tube reaction using alternative isothermal enzymes.
Key features:
- Extreme thermostability* ( < 98ºC )
- Superior strand-displacement activity
- 65ºC temperature optimum
- 5’-3’ exonuclease minus
- 3’-5’ exonuclease minus
*no loss of activity is observed in a LAMP reaction following 5 minutes @ 95ºC step.
Advantages of HD-LAMP:
- Template denaturation step at 95ºC prior to amplification has been shown to increase target sensitivity[1]
- Increased primer specificity – reduces false-priming that may occur at lower temperatures.
- Simple template extraction (cell-lysis).
- Sample heat-lysis, amplification and detection in a single tube.
[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 |
| TIN-001 | 5000u Thermodesulfatator indicus (exo-) LF DNA polymerase @ 8u/µl |
| TIN-002 | 25000u Thermodesulfatator indicus (exo-) LF DNA polymerase @ 8u/µl8000u |
| TIN-002HC | 25000u Thermodesulfatator indicus (exo-) LF DNA polymerase @ 100u/µl |
| iBuffer4-001 | 2 x 1.5ml iBuffer 4 |
| iBuffer4-002 | 15ml iBuffer 4 |
Effect of heat-denaturation on Tin(exo-) LAMP (Human gDNA template): Time-to-threshold comparison:
25µl HD-LAMP reactions: 8U DNA Pol, 1X manufacturers supplied reaction buffer (‘Tin(exo-)’ – iBuffer4, ‘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. ‘Tin(exo-)’ 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.
Tin(exo-) vs competitor ‘N’ (LAMP Pol 3.0) and competitor ‘L’ (thermostable LAMP Pol)
