Table 1 compares classical Phi29 and EquiPhi29 DNA polymerases. ![]() For this reason, exo-resistant random primers are recommended. Moreover, it retains all the benefits of the wild-type enzyme, including high processivity (up to 70 kb), strong strand displacement activity, and 3′→5′ exonuclease (proofreading) activity. This enzyme is significantly superior over Phi29 in protein thermostability, reaction speed, product yield, and amplification bias. This property is used to synthesize DNA complementary to single-stranded DNA templates. The addition of mononucleotides from dNTPs to the 3-OH terminus of DNA is catalyzed. It lacks the 53 exonuclease activity of the native enzyme. Thermo Scientific offers an improved EquiPhi29 DNA polymerase, which is a proprietary Phi29 DNA polymerase mutant developed through in vitro protein evolution. KLEN-RO) is a DNA-dependent 53 polymerase with 35 exonuclease activity. Phi29 DNA polymerase is the main enzyme of choice for WGA. Various WGA techniques have been developed that differ both in their protocols and ease of use. DNA amplified by WGA is used in downstream next-generation sequencing, Sanger sequencing, genotyping with microarrays, and single nucleotide polymorphism (SNP) genotyping. PCR-based labeling is superior to random-primed labeling with Klenow fragment if template. Amplification of Klenow gene fragment on ice for 45 min, 5 M NaCl and 20 Triton X-100 were added to a final concentration of 100 mM and The Klenow gene fragment of E. ![]() This is particularly useful in genetic disease research, where many repetitions are required. To increase the amount of limited DNA targets, isothermal whole genome amplification (WGA) is the most efficient technique.
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