Talk:Reverse Transcription Loop-mediated Isothermal Amplification

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Molecular Biology: Genetics / MCB

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Suggest merging with article on LAMP

Might I suggest merging this with the page on Loop-mediated isothermal amplification? RT-LAMP is just LAMP with RT added, to allow it to more efficiently amplify from RNA templates. Even if you don't add the RT, LAMP will occasionally work on RNA templates as the Bst DNA pol has a little bit of RNA dependent DNA polymerase activity. The mechanism of RT-LAMP as described here is pretty much the same as LAMP for a DNA target. Which it is, once the RT has synthesized cDNA. As the schematic shows, RT-LAMP it has to be initiated by the correct primer (the BIP), followed by displacement by the B3; the other thing that can happen is RNAseH degradation of the RNA part of the RNA:cDNA hybrid (the commonly used AMV RT has endogenous RNAseH activity, which turns out to be useful in RT-LAMP, unlike RT-PCR). This is a little different from the straight-up DNA LAMP, where both the FIP and BIP (and the F3 and B3) take part in initiation.--RobertM75 (talk) 06:43, 24 January 2015 (UTC)[reply]

Seems not very neutral

This article does come across as very pro-RT-LAMP. It's a fine technique for detecting RNA viruses, but the "advantages & disadvantages" section, as well as the rest of it, comes across like an ad for RT-LAMP (which is funny because I don't think even Eiken makes much money off of RT-LAMP). Off the top of my head - a major disadvantage of RT-LAMP for detecting RNA viruses is not just that the primer design is intrinsically complex because of the crazy reaction scheme. The bigger problem in my mind for RNA viruses is that you have to find ~120 bases of conserved sequence (~160 bases if you want to use loop primers) subject to pretty tight constraints on spacing, Tm, etc. If you've ever looked at a sequence alignment for RNA viruses, this can be pretty hard to do - you may end up with primers that only work for a specific clade, or you may find a conserved target that turns out to be conserved across related viruses as well. By contrast PCR primers are a lot easier, in that forward + reverse + probe you have a much easier time finding 3 regions of ~20 bases, with more freedom on the spacing.--RobertM75 (talk) 06:43, 24 January 2015 (UTC)[reply]

The first schematic needs minor review

In the penultimate step (the one right before the last black arrow) in the first schematic, the third box of the displaced cDNA strand should be green, not brown. Furthermore, I think that "DNA Polymerase" in the second yellow ellipse should be changed to "Reverse Transcriptase" since RNA serves as a template for synthesis in this step. Given the fact that "Reverse Transcriptase" is also a "DNA Polymerase", these two types of enzymes should be specified more precisely (e.g. "DNA-dependent DNA Polymerase" instead of just "DNA Polymerase"). --Marko108~cswiki (talk) 21:53, 27 November 2017 (UTC)[reply]

S4155659@mkhb.moe.gov.sa

5sdeeedaqssssrdseedrrzedessezseseeeeeeff 2001:16A2:F291:4000:916B:A7B3:F59C:5366 (talk) 11:18, 22 February 2022 (UTC)[reply]