I am working on establishing a LAMP based protocol for detecting Yersinia enterocolitica- which is a food borne enteropathogn.

I designed the three primer pairs (F3, B3, FIP, BIP, LF, LB) from chromosomal beta-lactamse gene of Y.enterocolitica using Primer explorer (Eiken Chemical Co). I used warm start Bst polymerase (NEB), isothermal buffer, MgSo4 and dNTPs as per the recommended concentrations.And set up a 25ul reaction at 65 C for varying time intervals. My problem is that when I run my LAMP amplicons on agarose gels I can see bands even after 5 min, 10 min, 15 min, 20 min, 25min ,30 min, 40 min, 50 min and 60 min.These are sharp bands( just like PCR amplicons). But I cannot see any ladder like pattern on the gel. One thing more, my positive and negative controls wok fine.The designed primers are specific since they do not amplify DNA of E.coli or any other bacteria.

Would be grateful to the research gate members for valuable suggestions