ORA™ qPCR Green Dye Mix, 2X

£106.00£477.00

highQu qPCR mastermixes are based on the small molecular inhibitor technology Hot Start PCR allowing to achieve highest sensitivity and specificity under both standard and fast qPCR cycling conditions. They provide excellent results on both AT and GC rich templates and guaranty rapid extension with early Ct values with minimum or no optimisation.

Our mastermixes are supplied with PCR Water to guaranty the best performance. To suit the broad instrument range the ORA qPCR Green Mastermixes are available in different versions –with low or high ROX concentration.

 

£106.00
£477.00
£106.00
£477.00

Description

highQu ORA™ qPCR Green Dye Mix

Applications

  • 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

Benefits

  • Universal – both standard and fast cycling
  • Excellent for GC or AT rich templates
  • Highest sensitivity, rapid extension, early Ct values
  • 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.

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