I have carried out a LAMP reaction, and I have obtained the typical ladder pattern. I wanted to know whether I can cut one of those bands and sequence it, trying with the different primers I have included in the LAMP reaction. My aim is to carry out a BLAST search to confirm the specificity of this LAMP reaction.
LAMP is an isothermal amplification method designed to detect a target nucleic acid without requiring sophisticated equipment. LAMP provides high sensitivity (to fg or
Hi Aaron
I'm also working on LAMP assay, but we never tried to sequence the LAMP products since they are mainly concatamers (and not single amplicons as in PCR). However, it should be possible, at least in theory, and resulting sequences should be a "tandem" repeated target sequence. To test the specificity of the assay, we used Optigene (http://www.optigene.co.uk/) reagents (which include a fluorophore) for LAMP, and a Real Time PCR instrument to detect the fluorescent signals and to analyze the melting curves. Basically, you should compare the melting curves of your samples to that of the positive control: it is very easy, very precise and less time-consuming than sequencing. Hope this helps.
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