OptiGene/GspSSD Isothermal Mastermix (ISO-001)/GspSSD Isothermal Master Mix - 400 reactions/ISO-001

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OptiGene’s Isothermal Master Mixes offer unrivalled DNA amplification speedOptiGene’s Isothermal Master Mixes (ISO-001 and ISO-004) were compared to NEB’s WarmStart LAMP Kit (DNA & RNA)

A DNA target (VKORC1) was amplified from Human gDNA (Sigma). 25μl LAMP reactions were performed at 65ºC for 60 minutes on a real-time thermocycler (Roche LC-480) in duplicate.The ‘Time to result’ is defined as the time at which the amplification fluorescence value reaches a threshold of 10% of maximal fluorescence.

LAMP primers targeting the VKORC1 gene were designed using LAMPDesigner®.0.8uM FIP – CTGTTGCACCTACCCTTCTGGAGCAGGAGAGGGAAATATCA0.8uM BIP – GAAGTCAAGCAAGAGAAGACCTGAAAACTCCTGACCTCAAGTGA0.2uM F3 – ACCATTATTCTGTCTACCACAC0.2uM B3 – GACAGGGTTTCACCATGTT0.4uM LoopF – TCTCTTCCTCTGGCGTCT0.4uM LoopB – CTCACGCCTATAATCCTAGCATTOptiGene’s Isothermal Master Mixes (ISO-001 and ISO-004) show faster DNA detection when compared to the NEB Master Mix.

OptiGene’s Isothermal Master Mixes offer unrivalled RNA amplification speed

OptiGene’s Isothermal Master Mixes (ISO-001 and ISO-004) were compared to NEB’s WarmStart LAMP Kit (DNA & RNA) [Lot:0021609].ISO-001 and ISO-004 DO NOT contain a specific reverse-transcriptase (RT) enzyme.For a true comparison to the NEB mix (which contains a separate and specific reverse-transcriptase enzyme)0.5U AMV-RT was added to the OptiGene Master Mix reactions (ISO-001 inc. RT and ISO-004 inc. RT).Standard ISO-001 and ISO-004 RT-LAMP reactions were also compared to show the excellent inherent reverse-transcriptase activity of the mix.

A RNA target (Beta-actin) was amplified from Human Total RNA (Ambion).25μl RT-LAMP reactions were performed at 65ºC for 60 minutes on a real-time thermocycler (Roche LC-480) in duplicate.The ‘Time to result’ is defined as the time at which the amplification fluorescence value reaches a threshold of 10% of maximal fluorescence.

LAMP primers targeting the Beta-actin gene were designed using LAMPDesigner®.0.8uM FIP – GAGCCACACGCAGCTCATTGTACACGGCATCGTCACCAAC0.8uM BIP – CTGAACCCCAAGGCCAACCGGCTGGGGTGTTGAAGGTC0.2uM F3 – CCCTGAAGTACCCCATCGA0.2uM B3 – ACAGCCTGGATAGCAACGT0.4uM LoopF – GCCAGATTTTCTCCATGTCGTC0.4uM LoopB – CGAGAAGATGACCCAGATCATGT

OptiGene’s Isothermal Master Mixes (ISO-001 and ISO-004) show excellent speed in RT-LAMP.ISO-004 amplified 2X faster than the NEB mix, and almost 3X faster if a specificRT enzyme is added for a like-for-like comparison.

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