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Abstract: This protocol describes the preparation of reaction buffer for homemade LAMP / RT-LAMP reactions which involves the use of home-brewed BstLF and MMLV enzymes. It also contains the instructions to prepare the enzyme storage buffers. Home-brewed enzymes information: BstLF polymerase: expression and purification protocol. expression plasmid sequence. M-MLV reverse transcriptase: expression and purification protocol. expression plasmid sequence. Testing We test the buffers on SARS-CoV-2 LAMP or RT-LAMP reactions with synthetic dsDNA or ssRNA respectively. The general performance of Homemade buffer seems appropriate and no amplification was obtained for non-template controls (NTC). To properly formulate the buffer we tested different KCl concentrations which is known to be a critical factor in the buffer. For the home-brewed BstLF, we found that 50 mM KCl is the best concentration, displaying equal or better performance than the commercial buffer. No amplification was observed for 150 mM KCl formulation. [1] Zhang, Y. et al. Enhancing colorimetric loop-mediated isothermal amplification speed and sensitivity with guanidine chloride. Biotechniques69, (2020).