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You are here : PCR / DNA AmplificationPolymerases ╗ Pfu/Psp DNA Polymerase

Pfu/Psp DNA Polymerase

Pfu DNA polymerase generated PCR fragments exhibits the lowest error rate of any thermostable DNA polymerases. The fragments are blunt-ended for direct ligation

 
 
Cat.-no Description Amount Price € Shop
 S127  Pfu/Psp DNA Polymerase    1x250 units    60.00  add
 S117  Pfu/Psp DNA Polymerase    2x250 units   110.00  add
 S118  Pfu/Psp DNA Polymerase  10x250 units   550.00  add

Pfu/Psp DNA polymerase: Order/request by E-mail:

Pfu/Psp DNA Polymerase: Datasheet

Pfu/Psp DNA Polymerase: Deutsche Beschreibung



Features:
Pfu/Psp DNA polymerase replicates DNA at 75°C catalyzing the polymerization of nucleotides into duplex DNA in the 5´=>3´ direction in the presence of Mg+. Pfu DNA polymerase possesses 3' to 5' exonuclease proof reading activity that enables the polymerase to correct nucleotide-misincorporation errors. The enzyme has no 5'=>3' exonuclease activity.

Applications:
- blunt end PCR cloning
- PCR and primer extension where "high fidelity" is required
- Site-directed mutagenesis

Description: 
Pfu/Psp DNA polymerase, isolated from the archae bacteria Pyrococcus furiosus/species is a thermostable Polymerase of approximately 90000 daltons.  Base misinsertions that may occur during polymerization are rapidly excised by the proofreading activity of the polymerase. The Pfu DNA Polymerase has no detectable reverse transcriptase activity.

Concentration:
 5 u/µl

Unit definition:
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nM of dNTPs into acid insoluble material in 30 minutes at 75°C.

Storage Buffer:
50 mM Tris-HCl, pH 8.2, 0.1 mM EDTA, 0.1% Tween 20, 0.1% Nonident P40, 1 mM DTT, 50% Glycerol

Reaction Buffer 10 X:
100 mM KCl, 160 mM (NH4)2SO4, 20 mM MgSO4, 200 mM Tris-HCl, pH8.8, 1% Triton X-100, 1 mg/ml BSA

Quality control:
- Tested  for the DNA amplification of 2,2 kb from lambda DNA
- Contamination level check of bacterial DNA
- Purity by SDS-Page > 90 %

Usage:  
Standard protocol:
- Do not use dUTP or dITP or primers containing these nucleotides

Components Volume per reaction end conc.
 10X reaction buffer with MgSO4  5 µl  1X
 dNTP-Mix (40mM = 10mM each)  1.0 µl  200 µM each
 Up-stream primer (e.g. 20 µM)  0,5 µl  0.1-1.0 µM
 Down-stream primer (e.g. 20 µM)   0.5 µl  0.1-1.0 µM
 Template DNA (10 ng/µl)  1.0 µl   <= 0,5 µg
 Pfu/Psp DNA Polymerase (5 u/µl)  0.2 - 0,4 µl  1-2 units
 Sterile dest. Water (molecular grade)  up to 50 µl 
 

Note: 
- vortex all solutions carefully before using
- dispense all reagents on ice to avoid degradation of primers and dNTP's
- add the enzyme after Template DNA
- may you have to optimize the MgSO4 concentration for best result

General Thermo-Cycler protocol:

 Step  Time  Temperature
 Initial denaturation  1-3 min  95°C
 
25-35 Cycles:
 Denaturation
 Annealing
 Extension
 
 30-100 sec
 30-65 sec
 1-2 min (per 1kb)

 95°C
 37-69°C
 72-75°C 
 
 Final extension    5 min  72-75°C

Storage: at -20°C for 24 months

Transportation: on blue ice

Related products:



>>> Service pages in English <<< >>> Serviceseiten auf Deutsch <<<
Copyrightę GeneOn 2007-15 print page: Pfu/Psp DNA Polymerase
 


 
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Polymerases
 
one-Fusion high-speed-fidelity Polymerase
HighEnd and exclusive Polymerase: faster speed, more accuracy, better result
DFS Taq Polymerase DNA-free Discontinued
DFS Taq Polymerase (DNA free) is a highly purified polymerase (DNA-free) especially for PCR reactions where freedom from endogenous template (i.e., E. coli DNA) is essential
DFS-PLUS Taq Polymerase DNA-free
DFS PlusTaq Polymerase: more sensitivity; more processivity
H-SPlus-Taq DNA Polymerase
Hot Start Polymerase with ultra short inactivation time, free of MAB´s. The high concentrated versions are designed for Lyophilization
Hot-Start PCR: M-Superhot Taq Polymerase
Superior Taq Polymerase for Real Time PCR and Hot-Start PCR, low-copy number PCR, PCR of difficult templates, Hot-Start activity (only 5 min initial denaturation). The enzyme is developed to enhance the specificity, sensitivity and yield of DNA amplification. The enzyme provides a convenient setting up at room temperature
SNPase for SNP-Genotyping
Hot-Start Polymerase for SNP detection by allele-specific PCR and micro sequencing
Taq DNA Polymerase
Maximo Taq Polymerase is a highly purified polymerase for routine amplification
Taq DNA Polymerase 2X-preMix
Maximo Tag Polymerase as 2X-preMix includes all components (Taq DNA polymerase, dNTPs, MgCl2, and reaction buffer) in an optimal concentration for routine PCR
Taq DNA Polymerase blue ready-to-load
Maximo Tag-Blue Polymerase is a highly purified polymerase for routine amplification. The enzyme is ready-to-load. No extra dye or loading buffer is needed!
m-Anti-Taq
Enhancer for Polymerase reactions
p-Taq Polymerase
p-Taq Polymerase is developed to enhance the specificity and yield of DNA amplification. The polymerase provides a convenient setting up at room temperature because of blocking the the enzyme with optimized mix of inhibitors
Bst DNA Polymerase
Bst DNA allows sequencing through problematic secondary structures
Pfu/Psp DNA Polymerase
Pfu DNA polymerase generated PCR fragments exhibits the lowest error rate of any thermostable DNA polymerases. The fragments are blunt-ended for direct ligation
Pfu/Psp red DNA Polymerase RTL
Pfu DNA polymerase (RTU=READY-TO-LOAD) is a convenient Mixture of proof-reading Enzyme, red dye and loading buffer. It generated PCR fragments exhibits the lowest error rate of any thermostable DNA polymerases. The fragements are blunt-ended for direct ligation
Pfu/Psp DNA Polymerase 2X-preMix
Pfu DNA polymerase 2X-preMix is a convenient Mixture of proof-reading Enzyme, reaction buffer dNTP´s and MgCl2. It generated PCR fragments exhibits the lowest error rate of any thermostable DNA polymerases. The fragements are blunt-ended for direct ligation
Tth DNA Polymerase
Tth DNA Polymerase Maximo is used to reverse-transcribe RNA efficiently in the presence of manganese. PCR can be performed in the same tube using the intrinsic DNA polymerase activity simply by chelation of manganese cation and the addition of magnesium.



 
 
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