Bacillus subtilis phage phi29, recombinant, E. coli
For in vitro use only!
Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmol of dNTP into an acid-insoluble form in 30 minutes at 74 °C.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 6 months
Concentration: 10 units/μl
Phi29 DNA polymerase is a recombinant protein purified from E. coli cloned the gene encoding the DNA polymerase from Phi29 phage. Phi29 DNA polymerase is the replicative polymerase from the Bacillus subtilis phage Phi29 and possesses the highest processivity and strand-displacement activity among the known DNA polymerase.
Phi29 DNA polymerase contains a 3'→5' exonuclease activity that enables proofreading capability.
Phi29 DNA Polymerase (brown cap)
10 units/μl Phi29 DNA Polymerase in storage buffer
(25 mM NaH2PO4 pH 7.0, 150 mM NaCl, 125 mM Imidazole, 50 % Glycerol, 2.5 mM 2-Mercaptoethanol)
Reaction Buffer (purple cap)
10x conc. complete reaction buffer containing 500 mM Tris-HCl pH 7.5, 100 mM MgCl2, 100 mM (NH2)4SO4 and 40 mM DTT
|Phi29 DNA Polymerase||20 μl||100 μl|
|Reaction Buffer||100 μl||500 μl|
Recommended Assay Set-Up
|component||Cat. No.||stock conc.||final conc.||1 assay|
@ 20 μl
|PCR-grade Water||PCR-258||fill up to 20 μl|
|dNTP Mix||NU-1006||10 mM||125 μM||0.25 μl|
|Random Hexamers 1)||100 μM||5 μM||1 μl|
|Target DNA||1 μl|
|Phi29 DNA Polymerase||PCR-381|
|10 units/μl||5-10 units/assay||5-10 units|
1) The use of random primers (i.g. 5'-NNNN*N*N-3') with phosphothioate protection against 3' exonuclease activity is recommended.
Incubate at 30 °C.
Heat the mixture to 65 °C for 15 min.
Osama Alsmadi et al. (2009) Specific and complete human genome amplification with improved yield achieved by phi29 DNA polymerase and a novel primer at elevated temperature. BMC Research Notes. www.biomedcentral.com/1756-0500/2/48