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

Table. 2.

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

Multiplex-PCR PCR1 PCR2 PCR3
Allele Ppo-A1
Rht-B1b
Vrn-D1
TaCwi-A1b
Glu-A3ac
Lr34
Glu-A1c
Glu-B1b
KWSM003
TaSE96
PCR reaction components
Taq DNA polymerase (U) 2 4 4
Each dNTP (mmol/l) 3 4 3
Primer (μmol/l) 1/1/2z 0.5/1.5/1.5y 2/1/1/1x
Concentration of DNA (ng) 120 100 80
PCR amplification condition
C 94℃, 5 m 94℃, 5 m 94℃, 5 m
Denaturation 94℃, 1 m 94℃, 1 m 94℃, 1 m
Annealing 62℃, 1 m 62℃, 1 m 64℃, 1 m
Extension 72℃, 1 m 72℃, 1 m 72℃, 1 m
No. of cycles 35 33 33
Final extension 72℃, 10 m 72℃, 10 m 72℃, 10 m
Gel electrophoresis
Agarose (%) 1.5 1.5 1.5
Voltage (V) 135 135 135
Time (h) 1 1 1

zConcentrations of primers for Ppo-A1, Rht-B1b and Vrn-D1a, respectively.

yConcentrations of primers for TaCwi-A1b, Glu-A3ac and Lr34, respectively.

xConcentrations of primers for Glu-A1c, Glu-B1b, KWSM003 and TaSE96, respectively.

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