Sequences for the PCR Primers Used to Amplify SSR Loci in Soybean
These primers were developed by Perry Cregan (Soybean and Alfalfa Research Laboratory, USDA-ARS, Beltsville, MD) with extramural financial support from the United Soybean Board and the able technical assistance of Edward Fickus, Sarah Hyatt, Charles Quigley, Patrick Elia, Susan Fogarty, Jason Kenworthy, and Chris Lee.
PCR conditions for using these primers can be found here.
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PCR Reagents for Soybean SSR Amplification
1. 30 ng genomic soybean DNA
2. Buffer:
1. 50 mM KCl
2. 10 mM Tris-HCl (pH 9.0 at 25o C)
3. 0.1 % Triton X-100
3. 1.5 mM MgCl2
4. 0.15 mM for each of the NTPs
5. 1 unit Taq DNA Polymerase
Thermocycling Profile for Amplification of Soybean SSRs
1. 2 min at 95o C
2. 33 cycles of
1. Denaturation: 92o C
2. Annealing: opimum temperature or 47o C if not known
1. For better, but still specific amplification, 46o C will also work quite well
3. Extension: 68o C
Use equal times for denaturation, annealing, and extension. Time depends on PCR machine, volume of reaction, etc.
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