Three sets of primers, various conc. of Mg2+, 1x amp buffer (NEB), Bst2.0 warm start (NEB), 0.8M betaine, 1.4 mM each dNTPs, 25 ul finalreaction vulme, 63C, 1h, followed by 80C, 5 min. Sometimes we get expected ladder pattern of bands in AGE, next time using the same reagents, nothing. Have repeated several times- no gain.
hello sir, I am currently doing a research on LAMP technology and was wondering if i could ask you questions regarding LAMP primer designs ? your help will be much appreciated.
Hello Thungdemo Ovung!
The F3, B3, FIP, BIP, LF and LB primers were constructed using PrimerExplorer4 software for amplification of Pasteurella multocida B:2 gene (GenBank: AF016260). We basically followed the method: Nagamine K, Hase T, Notomi T (2002). Accelerated reaction by loop-mediated isothermal amplification using loop primers. Molecular and Cellular Probes. 16: 223-229.
hello Ajit,
Thanks very much for the Information. my research concerns mostly about the primer design as many people do not get their way around and getting a functional and validated primer.
Sir, i am sure you are an expert in the use of LAMP technology can you please hep me with my research the problems faced in the primer designs and reagents faced by many scientists/ your experience using LAMP technology.
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