phi29 DNA Polymerase

Description

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 30C. phi29 DNA Polymerase, 10X phi29 DNA Polymerase Reaction Buffer, and dNTP mixture are included. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back 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 30C for 2 hr. After the amplification reaction, electrophoresis was performed on a 0.7 % TAE agarose gel. M denotes an lambda HindIII ladder, and NTC denotes the no-template control. Back 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 30C 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 . Back 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 30C. An additional reaction using Competitor Cs mutant phi29 polymerase was incubated at 42C.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 lambda 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.