Incubation for 30 minutes at 37✬ in a total reaction volume ofĪll preparations are assayed for contaminating endonuclease and 5'-exonuclease activities. 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 DNA polymerase and a 3j 5j exonuclease, but does not have 5j 3j exonuclease activity. Used for 3'-end labeling and filling in 5'-protruding sticky ends (4)ġ0X DNA Polymerase I (Klenow) Reaction Buffer Unit Definition MFCD00283091 Storage: -20C UNSPSC Code: 12352204 General description: DNA polymerase I yields two fragments (small and large) upon protease digestion. DNA Polymerase I Large (Klenow) Fragment consists of a single polypeptide chain (68kDa) that lacks the 5->3 exonuclease activity of intact E. Primer Oligonucleotide 3j Template DNA 5j Klenow fragment 3j.0.5 and that the Klenow fragment is not precipitated quan-. E013 Description, DNA Polymerase I Large (Klenow) Fragment is the large fragment of E. Suitable for second strand cDNA synthesis (3) over, the techniques we have described for the cloning and expression of a gene fragment. Joyce (1984 BRL Focus 6 (1): 6) and del Solar and Espinosa (1991 NAR 19 (8) 1956) suggest that some restriction enzymes stay attached to the cut ends of molecules and may require phenol:chloroform extraction (the most frequent offenders are BamH I, Hind III and Pst I) prior to Klenow fragment fill-in.
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