How does the inner primer attached to the target gene in the first place, as there is no denaturation process in Loop‐mediated isothermal amplification (LAMP). Does the inner primer has the ability to divide the strand? How?
From the original LAMP paper (Notomi et al. 2000):
"Since hybridization of the four primers to the target DNA in the initial step was critical for the efficiency of LAMP, the sequences and sizes of the primers were chosen so that their melting temperatures (Tm) fell within certain ranges. The F2 and B2 sequences in FIP and BIP were chosen such that their Tm values fell between 60 and 65°C, the optimal temperature for Bst polymerase. "
So, while there is not complete melting of the target sequence, there is melting at the F1c priming site that allows for strand-displacement amplification by the polymerase.
Article Loop-Mediated Isothermal Amplification for Detection of Afri...
Albert Vill Hiba Riyadh Al-abodi so our target sequences are limited to sequences that have Tm values within certain ranges? what if we want to amplify target sequences that have no desired Tm values across or near the target sequence?
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