Products
Molecular Biology Reagents | Consumables
Nucleic Acid Amplification
PCR Series
2 x Fast sTaq PCR Master Mix (for PCR series)
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Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
2x Fast sTaq PCR Master Mix | G3304-01 | 1 mL |
G3304-05 | 5×1 mL |
Product Description/Introduction
This product is a ready-to-use PCR premix, containing modified Taq DNA polymerase, dNTPs and optimized PCR reaction buffer at a concentration of 2×, which is suitable for routine simple template PCR, colony (liquid) PCR. PCR amplification can be carried out by adding only the template, primers and ddH2O, so that the concentration of the solution is 1×. Strong amplification ability, fast amplification speed, extension speed up to 5-15 s/kb, amplification product 3' end with A base. Loading Dye has been added to the product, and the amplified product can be directly used for agarose gel electrophoresis detection.
Storage and Shipping Conditions
Ship with wet ice; Store at -20, valid for 12 months. Avoid repeated freezing and thawing.
Product Contents
Component | G3304-01 | G3304-05 |
2x Fast sTaq PCR Master Mix | 1 mL | 5×1 mL |
Manual | One copy | |
Assay Protocol / Procedures
1. Recommended PCR reaction system
Component | 20 μL rxn | 50 μL rxn | Final Concentration |
Template DNAa | 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 sTaq 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: 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 bacterial fluid or crude extract sample, the volume added to reaction is no more than 10% of total reaction volume. Higher amounts of template tend to lead to non-specific amplification. Lower amounts of template tend to lead to low PCR amplification efficiency.
b: The range of final primer concentration used is 0.2-1.0 μM，and 0.4 μM is recommended. Too little primer may result in low or no amplification yield, and too much primer may result in non-specific amplification.
c: For high GC content template, DMSO with no more than 10% total volume can be added.
2. Recommended PCR amplification conditions
Step | Temperature | Time | Cycles |
Initial Denaturationa | 95 | 30 s-120 s | 1 |
Denaturation | 95 | 5-10 s | 25-35 |
Annealingb | 50-72 | 10-30 s | |
Extensionc | 72 | 5-15 s/kb | |
Final extension | 72 | 5-10 min | 1 |
Hold | 4 | Forever |
a: A pre-denaturation time of 30 s is suitable for most conventional templates; complex templates can be denatured for up to 2 min.
b: For amplification of complex templates, containing concatenated primers, the annealing time can be extended up to 30 s.
c: The recommended extension step is 5-10 s/kb at 72° for simple templates such as plasmid,10-15 s/kb for routine genomic DNA templates, 15-30 s/kb for complex template.
Note
1. When using, please thaw and mix thoroughly, after completely thawed can be placed in 4 stable storage at least 2 weeks, avoid repeated freezing and thawing.
2. For your safty and health,please wear safety glasses, gloves, or protective clothing.
For Research Use Only!
