Incubation for 30 minutes at 37✬ in a total reaction volume ofĪll preparations are assayed for contaminating endonuclease and 5'-exonuclease activities. DNA polymerase I, the large fragment (Klenow fragment) is the protease hydrolysate of E. One unit is the amount of enzyme required to incorporate 10 nmoles of total nucleotide into acid insoluble form in 30 minutes at 37☌ Heat InactivationĠ.033 mM of each dCTP, dGTP, dTTP and dATP View the full answer Final answer Transcribed image text: DNA polymerase I and DNA polymerase III in E. First, the Klenow fragment does not incorporate non-natural nucleotides such as IndTP and 5-AITP opposite thymine even though they are arguably considered good mimetics of dATP (Supplemental Figure 3). ![]() coli encodes the subunit of DNA polymera. DNA polymerases, such as the Klenow fragment of DNA Polymerase I and T4 DNA Polymerase are often used to fill in (5 3) and chew back (3 5). It has distinct regions for binding single- and double-stranded DNA as well as domains for dNTP binding, polymerization, and 3-5 exonuclease ( 2 ).
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