Safe-Green 2X PCR Taq MasterMix 5.0 ml (200 Rxns)
Features and Benefits - Completely eliminating DNA staining and running dyes in both gel and electrophoresis buffer.
- The simplest PCR formulation on earth.
The Safe-Green
TM 2X PCR Taq MasterMix is a ready-to-use mixture of high quality Taq DNA
Polymerase, deoxynucleotides, and reaction buffer in a 2X concentration. It contains
all the necessary reagents for the amplification of DNA templates as well as all essential
elements required for downstream electrophoresis, completely eliminating the needs for
EB and other gel staining dyes which are of significant environment concerns. To set up a
PCR reaction: simply add DNA template, primers and ddH
2O. Up to 6 kb templates can be
amplified with a single base (A) overhang at the 3’-end of PCR products.
Shipping and Storage Keep at –20°C for long term storage and at 4°C for 3 months for daily use.
Protocol PCR reactions should be assembled in a nuclease-free environment. DNA sample
preparation, reaction mixture assemblage and the PCR process, in addition to the
subsequent reaction analysis, should be performed in separate areas. The use of “clean”,
automatic pipettors designated for PCR and aerosol resistant barrier tips are recommended.
Always keep the control DNA and other templates to be amplified isolated from the other
components.
A control reaction, omitting template DNA, should always be performed to confirm the
absence of contamination.
1. Add the following components to a sterile 0.2 ml PCR tube sitting on ice:
| Components | Volume | Final Concentration |
| Template DNA | 100 ng | 2 ng/µl |
| Forward primer (10 µM) | 1 - 2 µl | 0.2 - 0.4 µM |
| Reverse primer (10 µM) | 1 - 2 µl | 0.2 - 0.4 µM |
| Safe-GreenTM 2X PCR Taq MasterMix | 25 µl | 1X |
| Nuclease-free H2O | up to 50 µl | - |
Note: We recommend preparing a pre-mix for multiple reactions to minimize reagent loss and enable accurate pipetting. 2. Mix contents of tube and centrifuge briefly.
3. Incubate tube in a thermal cycler at 94°C for 3 mins to completely denature the
template.
4. Perform 30-40 cycles of PCR amplification as follows:
Denature: 94°C for 30 sec
Anneal: 45°C - 72°C for 30 sec
Extend: 72°C for 1 min/1 kb template
5. After additional elongation at 72°C for 5 mins, the amplified PCR products are ready for
direct gel electrophoresis (no running and staining dyes in both running buffer and gel!)
and to be stored at -20°C until use.
6. When performing gel electrophoresis, a specially formulated DNA markers have to be
used as there is no DNA staining dye in both gel and running buffer.
7. It is strongly recommended to use the following DNA markers to analyze the PCR product:
Safe-Green™ 100 bp Opti-DNA Marker (Cat. No. G473)
Safe-Green™ 1 kb Opti-DNA Marker (Cat. No. G474)
