Korean Journal of Breeding Science

Conditions of multiplex-PCR for identification of Korean white winter wheat cultivars

Multiplex-PCR	PCR1	PCR2
Allele	Pina-D1/Ppo-A1/ Vrn-D1a	Ppo-B1/Glu-B3h
PCR reaction components
Taq DNA polymerase (U)	4	4
Each dNTP (mmol/l)	3	3.5
Primer (μmol/l)	2/2/3z	2/2.5y
Concentration of DNA (ng)	100	100
PCR amplification condition
Pre-denaturation	94℃, 5 m	94℃, 5 m
Denaturation	94℃, 40 s	94℃, 1 m
Annealing	60℃, 40 s	60℃, 1 m
Extension	72℃, 1 m 30s	72℃, 1 m
No. of cycles	35	33
Final extension	72℃, 5 m	72℃, 10 m
Gel electrophoresis
Agarose (%)	1.5	1.5
Voltage (V)	135	135
Time (h)	1	1

^zConcentrations of primers for Pina-D1, Ppo-A1 and Vrn-D1a, respectively.

^yConcentrations of primers for Ppo-B1 and Glu-B3h, respectively.