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Figure 1. Two rapid molecular marker assays to distinguish between PI 361088B and wild-type GmFAD3A alleles. (A) Sequence of nucleotides 301 to 330 of the GmFAD3A coding sequence from ‘Williams 82’ and PI 361088B DNA. Amino acid sequence from 101 to 110 is displayed below their corresponding codons. The XmnI recognition site is indicated by a black bar above the sequence. Note that the XmnI recognition site is disrupted by the two nucleotide insertion in the PI 361088B sequence. (B) A 200-bp sequence encompassing the region shown in Fig. 1A was amplified from Williams 82 or PI 361088B genomic DNA and treated with or without XmnI endonuclease. Products were then resolved on a 2% agarose gel (shown). When XmnI cleaves the Williams 82 polymerase chain reaction (PCR) product, bands of 140 and 60 bp are generated. (C) Melting curve analysis on Williams 82 and PI 361088B PCR products that have been digested with XmnI. The melting curve average was 77.4°C for the Williams 82 product and 79.4°C for the PI 361088B product.
