2X PCR HotStart MasterMix with dye

Details for Product No. ABIN4219189,
Polymerase Chain Reaction (PCR), Multiplex Assay (MA)
Purpose Problems with non-specific amplification and primer-dimer formation? Try HotStart DNA Polymerase.
Specificity With HotStart DNA Polymerase, you can achieve
- The highest specificity with minimal background
- Superior performance
- Improved yield of desired product
Characteristics HotStart DNA Polymerase contains a proprietary antibody that blocks polymerase activity at low temperatures. During the initial denaturation step at 94 °C, the antibody dissociates from Taq DNA polymerase and restores enzyme activity. This feature significantly reduces non-specific product formations that would otherwise compete for reagent availability. Thus, HotStart DNA Polymerase offers improved yield of desired PCR products. Now HotStart DNA Polymerase comes in a 2X PCR MasterMix format which provides all ingredients necessary for PCR in a premixed and optimized format that simplifies the PCR workflow. Also the available 2X PCR HotStart MasterMix with dye includes, in addition, an inert green dye blend and a stabilizer which allow for direct loading of the PCR product(s) onto an agarose gel. The green dye blend resolves during gel electrophoresis into a turquoise band at ~4000bp and a yellow band at the ~50bp region. The resolving bands of dye allow easy visualization of the real time progress of your gel electrophoresis, where the yellow band indicates the migrating front in the gel.
Components 5 x 1.0 ml vials
Application Notes - Assays with prolonged reaction setup or liquid handling
- Multiplex PCR
- Specific amplification of difficult templates (i.e. G-C rich)
- Low copy PCR assays
- TA Cloning

All PCR experiments should be assembled in a nuclease-free environment. In addition, DNA sample preparation, reaction set-up, and subsequent reaction(s) should be performed in separate areas to avoid cross contamination. The use of "clean" pipettors designated for PCR and aerosol resistant barrier tips are recommended. A negative control reaction (omitting DNA template) should be performed in tandem with sample PCR to confirm the absence of DNA contamination.
1. Add the following components into a sterile 0.2 ml PCR tube on ice (50 μlreaction volume).
- 2X PCR MasterMix 25 μl (Final Concentration 1X)
- DNA Template ~100 ng (Final Concentration ~2 ng/μl)
- Forward Primer (10 μM) 1-2.5 μl (Final Concentration 200-500 nM)
- Reverse Primer (10 μM) 1-2.5 μl (Final Concentration 200-500 nM)
- Nuclease-free H2O Up to 50 μl
2. Mix tube contents carefully and thoroughly, then collect all reagents by a brief centrifugation.
3. Choose an appropriate PCR amplification protocol for different DNA Polymerases.
4. Final PCR products are analyzed by agarose gel electrophoresis

Restrictions For Research Use only
Concentration 2 x
Storage -20 °C
Storage Comment Store at -20°C.