highQu ORA™ qPCR Green Dye Mix
- Dye based qPCR on instruments calibrated with low or high ROX concentration available.
- qPCR from gDNA, cDNA, viral DNA, low copy number genes
- Relative gene expression analysis, absolute quantification
- Universal – both standard and fast cycling
- Excellent for GC or AT rich templates
- Highest sensitivity, rapid extension, early Ct values
- Supplied with PCR Water
- Use special primer selection programs for good planning.
- Work with amplicons in a range of 80-200, max 400 bp.
- Take typical measures to prevent PCR cross over contamination, keep your bench clean, wear gloves, use sterile tubes and filter pipet tips.
- Run reactions in triplets; include a no-template control and positive control in parallel.
- Thaw and keep reagents on ice. Mix well before use.
Prepare a 20 µl reaction:
Reverse Primer 100 – 400 nM final c.
Forward Primer 100 – 400 nM final c.
cDNA Template or gDNA Template <100ng or <1 μg
PCR Water to 10 μl
ORA™ qPCR Mix, 2X 10 μl
Mix gently, avoid bubbles
Place into the instrument (SYBR® Green or FAM channel), set like:
Initial denaturation 1 cycle: 95°C – 2 min for cDNA, or
1 cycle: 95°C – 3 min for gDNA
Denaturation 40 cycles: 95°C – 5 sec
Annealing/extension 40 cycles: 60-65°C – 20-30 sec
Follow instrument instructions for melt curve analysis