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. To reduce the risk of
contamination, pipetting errors and to increase the repeatable of results the 2X-preMix contains an optimized mixture of enzyme, dNTP's and reaction buffer. Just add your template DNA and
primers.
Applications:
- blunt end PCR cloning
- PCR and primer extension where "high fidelity" is required
- Site-directed mutagenesis
- PCR where visual control is needed
Description:
Pfu/Psp DNA polymerase 2X-preMix is isolated from the archae bacteria Pyrococcus f-species, 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/Psp DNA Polymerase has
no detectable reverse transcriptase activity.
Concentration: Premix 2X (25µl per reaction)
Unit definition:
One unit is defined as the amount of enzyme required to catalyze the incorporationof 10 nM of dNTPs into acid insoluble material in 30 minutes at 75°C.
Storage: at -20°C for 24 months
Transportation: on blue ice
- Tested for the DNA amplification of 2,2 kb from lambda DNA
- Contamination level check of bacterial DNA
- Purity by SDS-Page > 90 %
- Do not use dUTP or dITP or primers containing these nucleotides
Components | Volume per reaction | end conc. |
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 2X-preMix | 25 µl | 1 unit |
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 Mg+ concentration for best result
General Thermo-Cycler protocol:
Step | Time | Temperature |
Initial denaturation | 1-3 min | 95°C |
|
|
95°C |
Final extension | 5 min | 72-75°C |
Loading on the gel:
Recommended volume is 10 µl of reaction mixture
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
* availibility of sample size may be limited
Final price excl. shipping costs3
Dear customer, GeneON likes to send free samples to convince the valued customer about the quality. Please understand that the shipping costs may be very high to some destinations. That is the reason why we cannot assure to fulfil all sample requests. Sorry for that, please understand. You may also set an inquiry/order directly by e-mail .