Figure 2
Amplification and standard curves of USS-sbLAMP reactions for detection of SNV p.H1047R utilising a synthetic plasmid DNA template containing a 531 bp fragment of the gene region. (a) Amplification curves of the WT template at different DNA concentrations ranging from 107 to 103 copies/reaction (31.356 pg/reaction to 0.003 pg/reaction respectively) using the WT reaction. (b) Amplification curves of the MT template at different DNA concentrations ranging from 107 to 103 copies/reaction (31.356 pg/reaction to 0.003 pg/reaction respectively) using the MT reaction. (c) Standard curve of the DNA template harbouring the WT allele, showing linear fitting equation and coefficient of determination. (d) Standard curve of the DNA template harbouring the MT allele, showing linear fitting equation and coefficient of determination. (e) Standard curve of the DNA template harbouring the WT allele with the addition of background Salmon testes sonicated DNA at 13 ng/reaction (+S). Results from the specific amplification of mixed populations are depicted in dark pink and labelled as MPWT reaction+S. (f) Standard curve of the DNA template harbouring the MT allele with the addition of background Salmon testes sonicated DNA at 13 ng/reaction (+S). Results from the specific amplification of mixed populations are depicted in dark pink and labelled as MPMT reaction+S. Error bars are displayed at one standard deviation. Reactions were considered as negative at time-to-positive (TTP) above 25 min. No template control (NTC) was included. Average of two experiments performed in triplicates.
