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4. 2 x Fast Pfus PCR Master Mix, 5 mL $350
$350.00
In Stock
Description
Product Information
| Product Name | Cat.No. | Spec. |
| 2 x Fast Pfus PCR Master Mix | G3305-01 | 1 mL |
| G3305-05 | 5×1 mL |
Description
This Fast Pfus PCR Master Mix is a 2×concantrated solution of optimized Pfu DNA polymerase, dNTPs and all other components required for PCR, expeceted DNA template and primers. This pre-mixed formulation saves time and reduces contamination due to a reduced number of pipetting steps during PCR set up. The optimized Pfu DNA polymerase ensure effieicnt PCR reaction, high fidelity, and high specificity, and with quick annealing speed of 5-15s/kb. 2×Fast sTaq PCR Master Mix is premixed with DNA loading dye, so the obtained product can ber direct loading on gel.
Storage and Handling Conditions
Transport use wet ice. Store at 4℃ for periods up to 12 months. For longer periods, stored at -20℃.
Assay Protocol
1.Gently vortex and briefly centrifuge PCR Master Mix(2×) after thawing.
2.Place a thin-walled PCR tube on ice and add the following components for each reaction accoridng to your desired reaction volume.
| Component | 20 μL rxn | 50 μL rxn | Final Concentration |
| Templatea | Variable | Variable | as required |
| Forward Primer (10 μM)b | 0.8 μL | 2 μL | 0.4 μM |
| Reverse Primer (10 μM)b | 0.8 μL | 2 μL | 0.4 μM |
| 2×Fast Pfus PCR Master Mix | 10 μL | 25 μL | 1× |
| (DMSO,optional)c | (0.6 μL) | (1.5 μL) | (3%) |
| ddH2O | Add to 20 μL | Add to 50 μL |
a:template quality is important for PCR reaction. Too little template result in low efficiency of PCR reaction, and excess template can result in nonspecific amplification. If the template is plasmid or from bacteriophage, 50ng-5pg added in 50 μL rnx is recommended. If tempalted is genomic DNA, 250ng-50ng added in 50 μL rnx is recommended. If template is cDNA, dilute your original cDNA by 2-100 times, and the volume of diluted cDNA added to reaction is no more than 10% of total reaction volume. If templtae is culture liquid, the volume added to reaction is no more than 10% of total reaction volume.
b:the appropriate primer concentration is between 0.2-1.0 μM，and 0.4 μM is recommended.
c: for high content GC template, DMSO with no more than 10% total volume can be added to improve PCR efficiency.
1.Gently vortex the samples and spin down.
2.When using a thermal cycler that does not contain a heated lid, overlay the reaction mixture with 20 μL mirenal oil.
3.Perform PCR using the recommened thermal cycling conditions below:
| Step | Temperature | Time | Cycles |
| Initial Denaturationa | 98℃ | 30 s-120 s | 1 |
| Denaturation | 98℃ | 5-10 s | 25-35 |
| Annealingb | 50-72℃ | 10-30 s | |
| Extension | 72℃ | 5-15 s/kb | |
| Final extension | 72℃ | 5-10 min | 1 |
| Hold | 4-16℃ | Forever |
a: in the initial denaturation step, 30s is suitable for most common template. For complex template, the time can be extened to 2 min.
b: in the Annealing step, the time 30s is suitable for complex template.
Note:
For your safety and health, please wear lab coat and disposable gloves.
For Research Use Only!
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