Takara phi29 DNA Polymerase. 250 U

Isothermal DNA polymerase derived fromBacillus subtilisphage phi29 with strong strand displacement and 3’-5’ exonuclease activities, enabling high-fidelity amplification of long DNA templates at 30℃. phi29 DNA Polymerase, 10X phi29 DNA Polymerase Reaction Buffer, and dNTP mixture are included.

phi29 DNA Polymerase successfully amplifies varying types of DNA templates across a wide input range. Amplification reactions were performed using phi29 DNA Polymerase (Cat. # RR340A) and 3 Phosphorothioate-modified Random Hexamer (Cat. # 3807) with HL60 genomic DNA (gDNA; top), M13 mp18 single-stranded DNA (ssDNA; middle), or pUC19 plasmid DNA (bottom) as templates. Input amounts of 5 ng, 0.5 ng, 50 pg, and 5 pg of each DNA template were added to the reaction solution, respectively, and incubated at 30°C for 2 hr. After the amplification reaction, electrophoresis was performed on a 0.7% TAE agarose gel. m denotes an λ HindIII ladder, and NTC denotes the no-template control.

Yield of phi29 DNA Polymerase reactions over time. Amplification reactions were performed using pUC19 plasmid DNA as a template with phi29 DNA Polymerase and 3 Phosphorothioate-modified Random Hexamer (n = 2). 0.5 ng of pUC19 plasmid DNA was added and the reaction was incubated at 30°C for 2, 4, 6, 8, or 16 hr. At each timepoint, the DNA yield per reaction was measured. The results show that although sufficient yield was obtained after 2 hr, the reaction time can be extended for higher yields (4 to 16 hr).

phi29 DNA Polymerase from Takara Bio outperforms competitor products in terms of sensitivity and yield. Amplification reactions were performed using phi29 DNA Polymerase from Takara Bio or phi29 DNA polymerase from competitor companies, with pUC19 plasmid DNA (5 ng, 0.5 ng, 50 pg, or 5 pg). Each reaction was incubated at 30°C. An additional reaction using Competitor Cs mutant phi29 polymerase was incubated at 42°C.Panel A. Each reaction was incubated for 2 hr. Electrophoresis was performed on 0.7% TAE agarose gels to assess for DNA amplification. m denotes a λ Hind III ladder, and NTC denotes the no-template control.Panel B. The DNA yield for all the reactions using 0.5 ng of template was measured at 2 and 4 hr. The phi29 DNA Polymerase from Takara Bio yielded the highest amount of DNA yield compared to other phi29 DNA polymerases from competitors. The yield was also comparable to that of the mutant phi29 DNA polymerase from Competitor C. No amplification was observed in the NTC reaction with Takara Bios enzyme.