For the procedure, I prepared the LAMP reagent mix with 1x isothermal buffer, 6 mM MgSO4, 1.4 mM dNTPs, and 9 µL nuclease-free water. I then added a primer mix consisting of FIP and BIP at 1.6 µM each, F3 and B3 at 0.2 µM each, and FL and BL at 0.4 µM each. After preparation, I heated the reaction mix at 95°C for 5 minutes. Finally, I added BST 3.0 polymerase and the template to the mix
* i used SYBR Green for the fluorescent dye
Hi,
It looks like no amplification. Pls check the primer sequences. Then, Pls check the amplification temperature. In general, the range is between 60-65 degree for LAMP PCR reaction. Please ensure to add one known Positive and one known Negative, then a no template control (to eliminate contamination) , run a gradient on the give range. I hope, it will help you to figure out the optimal temperature which required for specific amplification.
all the best
Kalyanaraman
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