2×Fast Pfus PCR Master Mix (1 mL)
- Stock: In Stock
- Model: 0437
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 ispremixed 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 | 20μL rxn | 50μL rxn | Final Concentration |
Templatea | Variable | Variable | as required |
Forward Primer (10μM)b | 0.8μL | 2μL | 0.4μM |
Reverse Primer (10μM)b | 0.8μL | 2μL | 0.4μM |
2×FastPfus PCR Master Mix | 10μL | 25μL | 1× |
(DMSO,optional)c | (0.6μL) | (1.5μL) | (3%) |
ddH2O | Add to 20μL | Add to 50μL |
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 50μL rnxis recommended. If tempalted is genomic DNA, 250ng-50ng added in 50μL rnxis 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 0.2-1.0μM，and 0.4μM 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 20μL mirenal oil.
3.Perform PCR using the recommened thermal cycling conditions below:
Step | Temperature | Time | Cycles | ||
Initial Denaturationa | 98℃
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New
DescriptionThis product 2×In-Fusion Cloning Mix Plus is an upgraded product of 2×In-Fusion Cloning Mix (G3350). 2×In-Fusion Cloning Mix Plus is compat..
$100.00
Ex Tax:$100.00
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