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. 2012 Mar 16;7(3):e32866. doi: 10.1371/journal.pone.0032866

Figure 7. Genotyping over a broad range of DNA amounts.

Figure 7

Melt-MAMA genotyping accuracy is not diminished at lower amounts of DNA, even at near-single copy for some assays. The sensitivity of individual melt-MAMAs varies greatly. This B. anthracis melt-MAMA (A.Br.003 clade) [4] accurately genotyped DNA regardless of starting amounts as long as it was sufficient to support amplification. (A & B) The respective amplification plots of genomic DNA of ‘G’ allele and ‘A’ SNP allele templates show the amplification curves of templates titrated in ten-fold serial dilutions and in replicates of eight. The number assigned to each amplification curve (1–8) denotes the DNA amount for the starting template. (C) The temperature-dissociation (melt) curve derivatives for all initial template amounts are shown (numbers denote DNA amount shown). This panel illustrates that genotyping accuracy was not affected by DNA amounts, even at near-single copy levels. Similar to TaqMan assays, the detection of low levels of DNA template by Melt-MAMA is also subject to stochastic sampling effects (B. anthracis single copy ∼6 fg), which is predictable using a Poisson distribution [4].