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OptiGene/Tin(exo-) LF DNA Polymerase/2 x 1.5ml iBuffer 4//iBuffer4-001

Tin(exo-) LF DNA Polymerase

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 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
TIN-001	5000u	Thermodesulfatator indicus (exo-) LF DNA polymerase @ 8u/µl
TIN-002	25000u	Thermodesulfatator indicus (exo-) LF DNA polymerase @ 8u/µl
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:Tin(exo-) vs competitor ‘N’ (LAMP Pol 3.0) and competitor ‘L’ (thermostable LAMP Pol)

tin poly

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.

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OptiGene/Tin(exo-) LF DNA Polymerase/2 x 1.5ml iBuffer 4//iBuffer4-001