Hi there, my lab recently opted to use LavaLAMP from Lucigen instead of the Warmstart kit from NEB. However, Lucigen has yet to create a colormetric kit for RT-LAMP, and adding phenol-Red (pH sensitive dye) and buffering to pH 8 via NAOH prior to amplification does not produce a colour change. I've also tested adding some acetic acid to the mixture and it clearly produces a colour change so there shouldn't be any issue with the dye. I ran the product on a gel and amplification clearly occurred, so the pH change was probably not sufficient to cause a colour change. My guess is that the mastermix may contain a buffer, which is preventing the pH from changing. I'm planning to do an experiment with minimal reagents, where I omit the mastermix, but I wanted to know if anyone else has tried this or has any suggestions?
Thank you!
Update to this question, I contacted Lucigen customer support. There is in fact a buffer mixture in both the enzyme solution as well as the 2x buffer. It is therefore incompatible with pH sensitive dyes. However there exists pH independent colorimetric methods such as HNB dye which relies on Mg+ ions, which should work well for colorimetric detection of amplification.
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