Nam risus ante, dapibus a molestie consequat, ultrices ac magna. Nam lacinia pulvinar tortor nec facilisis.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Fusce dui lectus, congue vel laoreet ac, dictum vitae odio. Nam lacinia pulvinar tortor sectetur adipiscing elit. Pellentesque dapibus sectetur adipiscing elit. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Fragments labeled at both 3 termini are used as the following: molecular-weight standards in Southern blotting (see Box 2). coli genomic DNA using oligonucleotide primers corresponding to the 16S rRNA locus.Sectetur adipiscing elit.
coli contamination is evaluated using 5 µL replicate samples of enzyme solution that are denatured and screened in a TaqMan qPCR assay for the presence of contaminating E. Single-stranded exonuclease is determined in a 50 µL reaction containing a radiolabeled single-stranded DNA substrate and 10 µL of enzyme solution incubated for 4 hours at 37☌.ĭouble-stranded exonuclease activity is determined in a 50 µL reaction containing a radiolabeled double-stranded DNA substrate and 10 µL of enzyme solution incubated for 4 hours at 37☌.ĭouble-stranded endonuclease activity is determined in a 50 µL reaction containing 0.5 µg of plasmid DNA and 10 µL of enzyme solution incubated for 4 hours at 37☌.Į. Purity is assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the band's mass corresponding to the protein of interest in the diluted sample. Physical purity is evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver-stain detection. Protein concentration is determined by OD 280 absorbance. Reactions were incubated for 10 minutes at 37☌, placed on ice and analyzed using the method of Sambrook and Russel (Molecular Cloning, v3, 2001, pp. Dilutions of the enzyme were made in a 50% glycerol Klenow (3ʹ→5ʹ exo–) storage solution and added to 50 µL reactions containing calf thymus DNA, 1x Klenow Reaction Buffer, 3H-dTTP and 100 µM dNTPs.
Klenow fragment molecular weight serial#
Unit activity was measured using a two-fold serial dilution method. Stop the reaction by adding EDTA (final concentration of 10mM) and heating at 75☌ for 20 minutes. Molecular Weight (g/mol) 68: Concentration: 5 U/L: Components: 10X Reaction Buffer: 500mM Tris HCl (pH 7.2 at 25☌), 100mM MgSO4, 1mM DTT: pH: 7. Incubate the reaction mixture at 25☌ for 15 minutes.ģ. DNA Polymerase I Large (Klenow) Fragment Shop Promega Klenow Fragment, DNA Polymerase I at Fisher Scientific Fisher Healthcare Fisher Science Education Sign Up.
Add dNTPs (each at a final concentration of 33 µM).
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