文字方塊: Pfu DNA polymerase ampilify the severial high yield DNA fragements og PCR products

 

 

 

 

 

 

 

Pfu DNA polymerase is a thermostable enzyme isolated from Pyrococcus furiousus. The enzyme replicates DNA at 75C, catalyzing the polymerization of nucleotides into duplex DNA in the 5'-3' direction. Pfu DNA polymerase possesses 3'-5' exonuclease (proofreading) activity. Base misinsertions that may occur during polymerization are rapidly excised by the proofreading activity of the polymerase.

Pfu DNA polymerase is recommended for use in PCR and primer extension reactions that require high-fidelity synthesis.

Pfu DNA polymerase-generated PCR fragments are blunt-ended.

Reaction Buffer (10x) with MgSO4:

200 mM TrisHCl (pH 8.8 at 25C), 100 mM KCl,

100 mM (NH4)2SO4,

20 mM MgSO4, 1.0% Triton X-100

Storage conditions: -20oC.

Procotol

Step

Temperature

Time

Cycle

Initial denaturation

94-95oC

1-3 min

1

Denaturation

94-95oC

0.5-2

25-35

Annealing

37-70

0.5-2

25-35

Extension

70-75

2min / 1kb

25-35

Final extension

70-75

5

1

 

 

 

Comparison of DNA polymerase and Pfu DNA polymerase

Pfu DNA polymerase

500U/100ul (5unit/ul)

 

Feature/characteristic

DNA polymerase

Pfu DNA polymerase

5‘-3’ polymerase

yes

yes

5‘-3’ exonuclease

No

yes

dNTP incorporate rate/sec

35 - 100

-

Error rate

2.4 x 10-5

0.4 x 10-5

Thermostability at 95oC

40 min > 50%

240 min > 70%

longest amplify

5 kb

10 kb

additional A tail

yes

No

application

PCR, RT-PCR

High fidelity, long PCR

 

 

 

 

 

文字方塊: Product NO.
specification
 
BL-T806
Pfu DNA Polymerase
500U/100ul
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 

 

 

 
   
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