Description
highQu ORA™ qPCR Probe Mix, 2X
Applications
- qPCR assays based on specific probes: TaqMan®, Molecular Beacons, Scorpions™ Probes
- Gene expression analysis, quantification of: gDNA, cDNA, viral DNA, low copy number genes
Benefits
- Universal – both standard and fast cycling, all probe qPCR assays
- qPCR of GC or AT rich templates, single-plex & multiplexing
- Rapid extension, early Ct
- Supplied with PCR Water
Protocol
Important Notes
- 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.
Specific Probe 200 nM final c.
cDNA Template or gDNA Template <100 ng or <1 μg
PCR Water to 10 μl
ORA™ qPCR Mix, 2X10 μl
- Mix gently, avoid bubbles.
- Place into the instrument 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
Anneal./extension 40 cycles: 60-65°C – 20-30 sec
- Follow instrument instructions for melt curve analysis
