The OmniAmp® polymerase is the only enzyme capable of Loop-mediated Isothermal Amplification (LAMP) from both RNA and DNA targets. The enzyme has a temperature optimum of 72°C, allowing increased specificity and flexibility compared to Bst Exo Minus or two-step processes. The native reverse transcriptase activity of this enzyme means you do not need to add a separate reverse transcriptase for RNA targets.
Flexibility: The enzyme comes with 10X OmniAmp Buffer, Magnesium Sulfate, and Betaine. The buffer is designed to allow for a wide range of additives and easy optimization following the instructions provided in the user manual.
Reliability: Achieve faster amplification at higher temperatures without sacrificing sensitivity. The enzyme has high target specificity and low background. Every batch is quality tested to ensure no unwanted nuclease activity will interfere with your sensitive applications.
Primer design: A robust LAMP reaction depends on proper design of the amplification primers. Please follow this PDF guide to properly use the PrimerExplorer software. The use of six primers is optimal; reactions are significantly slower and detection is less sensitive if only four primers are used.
|Figure 1. Quantitation of LAMP Amplification from a DNA Target |
Triplicate 25 µL LAMP reactions were set up using a primer set for a S. aureus ClfA target gene and included varying target DNA dilutions as indicated plus a No Target Control (NTC). Reactions were set up as recommended by each manufacturer. OmniAmp® Reactions: 1X Reaction Buffer plus 4 mM MgSO4 (10 mM total), 0.8 mM dNTPs, 150 mM betaine, 0.5X primers, 1X Fiona Green and 0.5X OmniAmp Polymerase. Bst (NEB) Reactions: 1X Reaction Buffer, 6 mM MgSO4 total, 0.8 mM dNTPs, 1X Fiona Green, 0.5X primers and 8 U Bst Polymerase. OmniAmp reactions were incubated at 68°C and Bst reactions at 65°C in a Bio-Rad CFX96™ Real-Time instrument (separate runs) and fluorescence was monitored over 60 minutes. Average fluorescence and standard deviations for each set of triplicate reactions are plotted.
|Figure 2. Quantitation of LAMP Amplification from a RNA Target. |
Triplicate 25 µL LAMP reactions were set up using a primer set for the Positive Control (MS2 RNA) included with the OmniAMP® Kit and 1 µL of Target RNA diluted as indicated plus a No Target Control (NTC). Reactions were set up as recommended by each manufacturer. OmniAmp Reactions: 1X Reaction Buffer plus 4 mM MgSO4 (10 mM total), 0.8 mM dNTPs, 150 mM betaine, 1X primers, 1X Fiona Green and 1X OmniAmp Polymerase. Bst 3.0 (NEB) Reactions: 1X Reaction Buffer, 6 mM MgSO4 total, 1.4 mM dNTPs, 1X Fiona Green, 1X primers and 8 U Bst 3.0 Polymerase. OmniAmp reactions were incubated at 70°C and Bst 3.0 reactions at 65°C in a Bio-Rad CFX96™ Real-Time instrument (separate runs) and fluorescence was monitored over 60 minutes. Average fluorescence and standard deviations for each set of triplicate reactions are plotted.
Licensing information: Lucigen is a fully licensed provider of LAMP reagents for research use. Patents WO 00/28082, WO 01/34790, and WO 01/77317 regarding the LAMP method are owned by the Eiken Chemical Co. Ltd. OmniAmp™ and Bst Polymerase, Exonuclease minus are sold by Lucigen under license for use in LAMP for research use only. The products may not be used for LAMP-based human or diagnostic purposes without obtaining a license from Eiken. US Patent 8093030 for OmniAmp DNA Polymerase is owned by Lucigen Corp.
It is the sole responsibility of the buyer to ensure that use of the product does not infringe the patent rights of third parties. If the purchaser is not willing to accept these use limitations, Lucigen Corporation is willing to accept return of the product for a full refund.
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Absence of Endonuclease or Nicking Activity: Incubation of OmniAmp™ DNA Polymerase with 1 µg of supercoiled pBR322 DNA for 16 hours at 70°C results in no detectable conversion to relaxed or linear forms detectable by agarose gel electrophoresis.
Absence of Exonuclease Activity: Incubation of of OmniAmp DNA Polymerase with 1 µg of HindIII-cut lambda DNA for 16 hours at 70°C resulted in no smearing of bands on agarose gels.
Purity: OmniAmp DNA Polymerase is >99% pure as determined by SDS-PAGE.
Exogenous DNA: There is no detectable DNA contamination.