Table 1

PCR conditions
PCR purpose PCR conditions (initial denaturation, number of cycles, denaturation, annealing, elongation, and a final extension step)
Enrichment for microsatellite-containing fragments 94°C for 2 min, followed by 12 cycles of 94°C for 15 sec, 55°C for 35 sec, 72°C for 90 sec
Recovery of microsatellite-containing fragments 35 cycles of 94°C for 15 sec, 55°C for 35 sec, 72°C for 30 sec, and a final extension at 72°C for 5 min
Preparation of products for cloning 94°C for 2 min, followed by 15 cycles of 94°C for 15 sec, 55°C for 35 sec, 72°C for 30 sec, and a final extension at 72°C for 5-10 min
Confirmation of cloned products 96°C for 2 min, followed by 33 cycles of 94°C for 40 sec, 57°C for 12 sec, 72°C for 30 sec, and a final extension at 72°C for 5 min
Genotyping with SSRs 96°C for 2 min, followed by 33 cycles at 94°C for 35 sec, annealing temperature* for 15 sec, 72°C for 30 sec, and a final extension at 72°C for 5 min

* Annealing temperatures for each SSR are shown in Additional file 1.

Rauscher and Simko

Rauscher and Simko BMC Plant Biology 2013 13:11   doi:10.1186/1471-2229-13-11

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