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2 X Fast Pfus PCR Master Mix Ready-to-use PCR Master Mix with Pfu DNA Polymerase DNTPs PCR Buffer

Volume:1ML
  • G3305-01

Product Description

Product introduction:

This ready-to-use PCR premix contains modified PFU DNA polymerase, DNTPs and optimized PCR reaction buffer at 2x concentration. It is suitable for conventional PCR, colony PCR, complex template and high GC template PCR. For PCR amplification, only template, primer and DDH2O were added to make the concentration of Mix solution 1X to carry out PCR reaction. It has strong amplification ability, fast amplification speed and extension speed up to 5-15s/ KB. It has high fidelity and high specificity. The amplification product is flat terminal. Loading Dye has been added to the products, and the amplified products can be directly used for agarose gel electrophoresis detection.   

 

 

Package Contents:

Cat. No.

Product description

Volume

G3305

2x Fast Pfus PCR Master Mix

1mL/5mL

 

Storage condition:

Wet ice bag transportation; Stored at -20℃ temperature, valid for 12 months.

 

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 Pfus 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 10 ng-1 pg per 50ul system; When using genomic DNA as template, the recommended addition amount is 250-50ng per 50ul system. When cDNA is the template, it is recommended to dilute it by 2-100 times and add no more than 10% of the total system. Too many templates will easily lead to nonspecific amplification, and too few templates will easily lead to low PCR amplification efficiency.  

a: The final primer concentration range is 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 nonspecific 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

98℃

30s-120s

1

Denaturation

98℃

5-10s

25-35

Annealingb

50-72℃

10-30s

Extensionc

72℃

5-15s/kb

Final extension

72℃

5-10min

1

Hold

4-16℃

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.

G3305-01

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