Korean Journal of Breeding Science :eISSN 2287-5174 / pISSN 0250-3360

Table. 3.

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.

Korean J. Breed. Sci. 2022;54:119-29 https://doi.org/10.9787/KJBS.2022.54.2.119
© 2022 Korean J. Breed. Sci.