This method of PCR amplification produces large concatamers of multi-looped amplicons those may show prominent visibility on the gel. But how this confers specificity of the amplification. Or if specificity is not the priority than what is?
Anirudha, to my understanding the purpose of LAMP, besides having great specificity, is that you can run a method similar to conventional PCR, but without needing a thermocycler. this is much cheap, because you can simply use a water bath and apart from these characteristics, depending on the agent you are trying to identify, you don´t need to perform a nucleic acid extraction. this saves money and time because you only incubate 1 hour at aprox 63°C. The idea of the technique is you can perform it in the field. To my experience I have performed it for CPV and It worked very well.
the specificity is very well expleined in Notomi et al., 2010.
cheers!
Sabrina is right.
LAMP does not require a thermal cycler and results can be visualized with naked eye without even running a gel. For some pathogens DNA extraction is no longer needed prior to LAMP. It takes less time, 30 min to 1 h is often enough. It is as sensitive as PCR if no more than PCR. Many reports found it 10 x sensitive than PCR. As far as specificity is concerned, you may do restriction enzyme digestion. The ladder banding pattern should be reduced to a few bands. Due to its sensitivity contamination might be an issue. So always run a negative control without DNA template to minimize false positive results.
I completely agree with Dr. Yao
you can visualize with naked eye your result! you can also try dying with sybr green,, you can read turbidity by spectophotometry. In my case I ussually use propidium iodide and to UV light you can see fluorescence for positive samples. I have attached some pics of a presentation for you to see how it looks like!
cheers!
If you want an isothermal that is as easy as PCR to design and even easier to run than LAMP, have you thought about Recombinase Polymerase Amplification (RPA)? It's the most PCR-like isothermal out there: http://www.twistdx.co.uk/our_technology/
RPA reactions typically take 5-15 minutes (so even faster than LAMP), you can use standard PCR primers (http://www.twistdx.co.uk/our_technology/pcr_primers_work_using_standard_rpa_reagents/), you can amplify several hundred bases, you can multiplex, good assays have single molecule sensitivity, it works with RNA and DNA and so on.
You don't even need a heater - you can just put the reactions under your arm and use body heat:
http://www.ncbi.nlm.nih.gov/pubmed/25372030
You can detect it colorimetrically like LAMP:
http://pubs.acs.org/doi/abs/10.1021/acssensors.5b00171?journalCode=ascefj
http://www.nature.com/articles/srep15028
http://pubs.rsc.org/en/content/articlelanding/2015/cc/c4cc10068a#!divAbstract
There are links to more publications (almost 100 now) here:
http://www.twistdx.co.uk/publications/
Best of all, it really works! There are some testimonials here: http://www.selectscience.net/suppliers/twistdx-limited/?compID=8753
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