STR locus

Primers [ ] (mM)

Reactive

Final [ ]

PCR conditions

Temp ˚C

Time

Cycles

Multiplex 1

DYS391

0.8

Buffer

1X

Pre-denaturalization

95

10 min

30

DYS437

0.7

MgCl2

1.75 mM

Denaturalization

94

1 min

DYS439

0.048

dNTP

200 mM

Annealing

55

1 min

DYS448

0.016

Betaina

0.83 M

Extension

70

1 min

DYS458

0.9

TaqPol

1 U

Post-extension

60

45 min

Multiplex 2

DYS19

1.2

Buffer

1X

Pre-denaturalization

95

10 min

30

DYS393

0.2

MgCl2

1.75 mM

Denaturalization

94

1 min

DYS438

0.25

dNTP

100 mM

Annealing

55

1 min

GATA-H4

0.4

Betaina

0.83 M

Extension

70

1 min

TaqPol

1 U

Post-extension

60

45 min

Multiplex 3

DYS389I

0.25

Buffer

1X

Pre-denaturalization

95

10 min

30

DYS389II

0.25

MgCl2

1.75 mM

Denaturalization

94

1 min

DYS390

0.2

dNTP

100 mM

Annealing

55

1 min

DYS392

0.5

Betaina

0.83 M

Extension

70

1 min

DYS456

0.2

TaqPol

1 U

Post-extension

60

45 min

DYS635

0.5