New England Biolabs, S0164S, Control LAMP Primer Mix (rActin)

Related Categories Isothermal Amplification & Strand Displacement,, PCR, qPCR & Amplification Technologies Applications Isothermal Amplification,, DNA Amplification, PCR & qPCR FAQ Q: In the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit, what is the target of the Internal Control Primer Mix (rActin)? A: The Internal Control LAMP Primer Mix targets actin RNA at an exon-exon junction. The primers also include a region that matches an ankyrin sequence in gDNA so low levels of amplification can be observed from mammalian gDNA but this is a minor contribution to overall signal given that rActin RNA is typically present at much higher copy numbers than gDNA. Q: My SARS-CoV-2 sample or Internal Control reaction turned orange/yellow upon sample addition. Does that mean that the sample contains nucleic acid? A: A color change to yellow or orange upon the addition of sample nucleic acid for the internal control or SARS-CoV-2 test sample reactions indicates that the input material is incompatible with the assay and the results are invalid. It should not be interpreted to signify the presence of target nucleic acid. Repeat with a lower sample volume or adjust the sample input pH to ~ 8. If the NTC or PC reactions immediately turn yellow or orange upon setup, repeat assay setup. Q: Does pre-heating the thermal block impact LAMP amplification reactions? A: Yes, we have observed that placing LAMP reactions on a pre-heated thermal block at 65 °C (rather than allowing the samples to warm to 65°C as the block heats up) tends to specifically improve detection of low input samples.