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Ready to use 2x Fast sTaq PCR Master Mix pcr kit
- Category: General Reagents >>>
- Supplier: Wuhan Servicebio Technology Co. Ltd.
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Product Overview
Description
Ready-to-use 2x Fast sTaq PCR Master Mix
Product Description
This ready-to-use PCR premix contains modified Taq DNA polymerase, DNTPs and optimized PCR reaction buffer. The concentration is 2X. It is suitable for conventional simple template PCR and colony (liquid) PCR. For PCR amplification, only template, primer and DDH2O were added, and the solution concentration was 1X to carry out PCR reaction. The amplification ability is strong, the amplification speed is fast, the extension speed can reach 5-15s/KB, the amplification product 3' end with "A" base. Loading Dye has been added to the products, and the amplified products can be directly used for agarose gel electrophoresis detection.
Product name | Cat. No. | Sepcification |
2x Fast sTaq PCR Master Mix | G3304-1 | 1ml |
G3304-2 | 5*1ml |
Product Usage
1. Recommended PCR reaction system:
Component | 20ul rxn | 50ul rxn | Final Concentraction |
Templatea | Variable | Variable | as required |
Forward Primer (10uM)b | 0.8ul | 2ul | 0.4uM |
Reverse Primer (10uM)b | 0.8ul | 2ul | 0.4uM |
2x Fast sTaq PCR Master Mix | 10ul | 25ul | 1x |
(DMSO, optional)c | (0.6ul) | (1.5ul) | (3%) |
ddH2O | Add to 20ul | Add to 50ul |
a: When using plasmid or phage DNA as template, the recommended dosage is 50ng-5pg per 50ul system; When using genomic DNA as template, the recommended addition amount is 250 ng to 50ng per 50ul system; When cDNA is used as template, it is recommended to dilute it by 2-100 times and add no more than 10% of the total volume. When using bacterial liquid or coarse sample as template, the addition amount should not exceed 10% of the reaction volume. Too many templates will easily lead to nonspecific amplification, and too few templates will easily lead to low PCR amplification efficiency.
b: The final primer concentration can be used in the range of 0.2-1.0um, and the recommended primer concentration is 0.4um. Too few primers may lead to low amplification yield or no amplification, and too many primers may lead to non-specific amplification.
c: High GC template can add no more than 10% additional DMSO to the reaction system.
2. Recommended PCR amplification conditions:
Step | Temperature | Time | Cycles |
Initial Denaturationa | 95℃ | 30s-120s | 1 |
Denaturation | 95℃ | 5-10s | 25-35 |
Annealingb | 50-72℃ | 10-30s | |
Extensionc | 72℃ | 5-15s/kb | |
Final extension | 72℃ | 5-10min | 1 |
Hold | 4℃ | Forever |
a: Predenaturation time 30s is suitable for most conventional templates, complex template can extend the denaturation time to 2min.
b: For complex templates, the annealing time can be extended to 30s with the amplification of annexed primers.
c: The extension speed of plasmid and other simple templates is recommended to be 5-10s/KB; Routine genome template recommended extension rate of 10-15s/KB; Complex templates 15-30S/KB.
Notes
1. For use, please defrost thoroughly and mix well. After complete melting, it can be stably stored at 4℃ for at least 2 weeks to avoid repeated freezing and thawing.
2. Please wear lab clothes and disposable gloves.
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