Parameters for SSR Screening and Primers in Brachypodium BAC End Sequences

Parameters for SSR screeding

SSR MotifMofif Length
Dinucleotide>= 12 nt
Trinucleotide>= 12 nt
Tetranucleotide>= 12 nt
Pentanucleotide>= 15 nt
Hexanucleotide>= 18 nt

Parameters for SSR flanking primer design

ParameterValue
Product size100-300, optimum: 150
Primer Tm50-50, optimum: 55, maximum Tm difference: 20
Primer GC%40%-60%
Self complementarity<= 8
3' self complementarity<= 3
Maximum # of N's0
Maximum polu X's5
Salt concentration50 nM
DNA concentration50 nM