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Home made qPCR Master Mix

ReagentVolume (for 150 µL)
qPCR mix (2×)100 µL
Sense primer (1 µM)2 µL
Antisense primer (1 µM)2 µL
Fluorescein (1 µM)2 µL
SYBR Green (40×) (Thermo Fisher Scientific S-7563)2 µL
Taq DNA polymerase:
JumpStart Taq Antibody (1:1) (New England Biolabs M0267X)
2 µL
Distilled water40 µL
Sterilize using a disposable 0.22-µm vacuum filtration system in a tissue-culture hood and store at room temperature.

qPCR Buffer (2×)

ReagentVolume/final concentration (for 1 mL)
PCR buffer (10×; 200 mM Tris-HCl,  500 mM KCl, pH 8.4)200 µL
MgCl225 mM
dNTPs10 mM
Sterilize using a disposable 0.22-µm vacuum filtration system in a tissue-culture hood and store at room temperature.

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