Magnesium was in the puffer, so we didn't change anything.
template LuxCDABEG is 6,3 kbp long.
The genome was rather control, because the plasmid from a recent midiprep should be purer (did also a proof-of-plasmid-transformation and they glowed).
Did you do an initial denaturation?
yes, like described in the instruction. https://www.neb.com/protocols/1/01/01/pcr-protocol-m0530
What's the program vs. amplicon length and primer melting temperatures?
It is written 15-30 seconds per kb , so we chose 180 seconds. 51.4 and 51.5 °C were the primer melting temperatures, and Stefan from the AEC said they always go a bit above the theoretical value to get a better result -> 52°C should work well.
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