The first few cycles of PCR are always the most important to successful amplification. Our mastermixes are never short of enough polymerase to kick start and then powerfully continue your amplification. MegaMix and MegaMix Blue are designed to cope with whatever is thrown at it. We incorporate two different Taq polymerases. The two Taqs complement each other and with the addition of Microzone’s own proprietary engineered PCR buffers, our mastermixes provide powerful, broad template amplification. PCR products generated are A-tailed and may be cloned into TA cloning vectors.
- Routine end point PCR
- Fast PCR
- Amplification of GC and AT rich templates
- Up to 6kb
- Methylated DNA
- Library Preparation
- TA Cloning
- More confidence in amplification and PCR test
- Mastermix format (Ready to use or 2x) for ease of use
- Both single and double concentrations. Get the right mix for your tests quickly and easily
- Easy set up and PCR optimisation
- Broad range of templates and conditions
MegaMix & MegaMix Blue
- As close as you’ll get to a one mix fits all.
- Exceptionally stable and can be freeze thawed on multiple occasions.
- High success rates up to 6kb.
- High yields under standard conditions.
- Provides effective and efficient amplification for GC and AT rich templates.
- Strong MgCl2 to support strong amplification.
- Thaw the MegaMix
- Add the desired volume (47ul in 50ul or if double then 25ul in a 50ul reaction volume)
- into the PCR tube.
- Add gDNA Template – 10 – 200ng. Less is always more.
- cDNA Template <100ng
- Add primers
- Final forward and reverse concentration of 0.2 – 0.6 µM from a 10 µM stock
- Overlay with mineral oil if necessary
- Place in a Thermal Cycler
Cycling Profile (guide only)
- Initial denaturation step: 95⁰C for 3 mins
- Then Cycle 25-30 times:
- Step 1: 95⁰C for 30 secs
- Step 2: Optimal annealing temp. of primers for 30 to 60 secs
- Step 3: 72⁰C for 45 to 60 secs
- Cool to room temperature
- Thaw reagents prior to use. Mix well prior to use.
- GC and complex templates may require higher denaturation temperatures
- Two step cycling protocols can increase speed. Following annealing just ramp the temperature back to denaturation.
- If using new primers or changing your mastermix it will be necessary to perform a new annealing temperature gradient. We suggest 1⁰C intervals and to start at 5⁰C below the calculated Tm of the primers and go to 5⁰C above the primer Tm.
- Primers design and quality have a large impact on the reaction. Taking time to design your primers using good techniques or software will benefit down the line. We do offer a technical note on primer design. Contact us to receive a copy.
We accept institutional purchase orders. Alternatively, purchases can be made with a debit or credit card through our UK distributor Clent Life Science