Pfu DNA polymerase
8-11 Munpyeongseo-ro, Daedeok-gu, Daejeon, 34302
Republic of Korea
Tel: +82-42-930-8777 (Korea : 1588-9788)
Fax: +82-42-930-8688 E-mail: [email protected]
[Cat. No.]
1301 Marina Village PKWY, Suite 110
Alameda, CA 94501, USA
Toll Free: +1-877-264-4300 Fax: +1-510-865-0350
E-mail: [email protected]
E-2015
[Lot. No.]
(V1/2016-02-15)
Korea Bio Park BLDG #B-702, 700 Daewangpangyo-ro
Bundang-gu, Seongnam-si, Gyeonggi-do, 13488
Republic of Korea
Tel: +82-31-628-0500 Fax: +82-31-628-0555
[Exp. date]
● Kit Content
Cat. No.
E-2015
E-2015-1
Pfu DNA polymerase
250 units
250 units
10X Reaction buffer
1 mL
1 mL
Dilution buffer
1 mL
1 mL
dNTP Mixture
1 mL
Note : For research use only. Not for use in diagnostic or therapeutic procedures. This product is not available for sale in the USA.
● Description :
Pfu DNA Polymerase is a thermostable DNA polymerase that catalyzes the polymerization of nucleotides into duplex DNA in the 5’ → 3’
direction in the presence of magnesium and exhibits 3’ → 5’ exonuclease (proofreading) activity. Bioneer’s Pfu DNA Polymerase is
recommended for use in PCR and primer extension reactions that required high fidelity.
● Applications : Polymerase Chain Reaction(PCR), Primer extension
● Supplied with Enzyme :
● Storage Condition :
- 10X Reaction Buffer (1 mL) : Tris-HCl, KCl, (NH4)2SO4,
Acetylated BSA, Triton X-100, 15 mM MgSO4, pH 9.0
- 10 mM dNTPs (1 mL)
50 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, Stablizers,
50 % Glycerol pH 8.2, store at –20℃
● Enzyme Concentration :
2.5 unit/μL
● Unit Definition :
One unit is defined at the amount of enzyme that will incorporates 10 nmol of dNTP into acid-insoluble material in 30 minutes at 72℃.
● Quality Assurance :
Nuclease activity is not detected after incubation of 1 μg of substrate DNA-supercoiled plasmid and Lambda/Hind III DNA - with 5 units of
Pfu DNA Polymerase in 50 μL reaction volume with the supplied Reaction buffer for 18 hr at 37℃ and 70℃.
● General Reaction Condition [50 μL reaction volume ]
- Reaction mixture
- PCR condition (Standard PCR)
Template*
variable
Step
Temp.
Time
Cycles
Primer (forward)
20 ~ 50 pmoles
Pre-denaturation
94℃
1 min
1 cycle
Primer (reverse)
20 ~ 50 pmoles
Denaturation
94℃
15 -20 sec
25-35
10X reaction buffer
5 μL
Annealing
AT℃
15 -30 sec
cycle
10mM dNTPs mix.
variable (1~5 μL)
Extension
72℃
1 min/kb
Pfu DNA Polymerase
2.5 units
Final extension
72℃
Optional. Normally
1 cycle
3-5 min.
D.W.
variable
Total
50 μL
* Amounts of template
(Plasmid and lambda DNA → more than 1 pg) (Bacterial genomic DNA → more than 100 pg) (Human genomic DNA → more than 1 ng)
● Note
- It is critical to without Pfu DNA polymerase until after the addition of dNTPs; Otherwise 3’ → 5’ exonuclease activity may degrade primers.
- Avoid multiple freeze-thaw cycles and exposure to frequent temperature changes.
● Ordering Information
[ Related BIONEER ‘s Pfu DNA Polymerase Products ]
Cat. No.
E-2016
D-3001
Products
Pfu DNA Polymerase 1,000 units, 10X Reaction buffer with MgSO4
10 mM dNTP mixture (1.0 mL), each dNTP 2.5mM
Consult
Research
Instruction
Use
For Use
Only
Catalog
Batch
Number
Code
Contains
Sufficient for
(n) tests
1
www.bioneer.com
Caution,
Consult
Accompanying
USE BY
Temperature
Limitation
documents
BQ-042-101-04
Revision : 4 (2015-11-06)