Tin(exo-) Isothermal Mastermix (ISO-001Tin)

OptiGene’s novel isothermal mastermix for amplification reactions requiring an initial heat-denaturing step.

Suitable for HD-LAMP (Heat-denaturing LAMP).

Contains the patented highly thermostable Tin(exo-) LF DNA Polymerase.

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 )

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 sensitivity¹

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.

¹Mori 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:	Description
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

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Effect of heat-denaturation on ISO-001Tin
LAMP (Human gDNA template): Time-to-threshold comparison:

ISO-001Tin vs competitor ‘N’ (LAMP Pol 3.0) and competitor ‘L’ (thermostable LAMP Pol)

iso001tin

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.