Pfu 2X Mastermix 5x100 rcs

Pfu/Psp 2X-preMix DNA Polymerase: Datasheet     
                                            
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 incorporation of 10 nM of dNTPs into acid insoluble material in 30 minutes at 75°C.

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

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:

Loading on the gel:
Recommended volume is 10 µl of reaction mixture

Storage: at -20°C for 24 months

Transportation: on blue ice