Crystal structures of open and closed forms of binary and ternary complexes of the large fragment of Thermus aquaticus DNA polymerase I: Structural basis for nucleotide incorporation. Structure of DNA polymerase I Klenow fragment bound to duplex DNA. DNA polymerase fidelity: Kinetics, structure, and checkpoints. Function and structure relationships in DNA polymerases.

Solid-state nanopore channels with DNA selectivity. Capture of a single molecule in a nanocavity Science 291, 636–640 (2001). Ion-beam sculpting at nanometre length scales. Evaluation of nanopores as candidates for electronic analyte detection. Fast DNA translocation through a solid-state nanopore. Unzipping kinetics of double-stranded DNA in a nanopore. Determination of RNA orientation during translocation through a biological nanopore. Sequence-specific detection of individual DNA strands using engineered nanopores. Recognizing a single base in an individual DNA strand: A step toward DNA sequencing in nanopores. Microsecond time-scale discrimination among polycytidylic acid, polyadenylic acid, and polyuridylic acid as homopolymers or as segments within single RNA molecules. Discrimination among individual Watson–Crick base pairs at the termini of single DNA hairpin molecules. Rapid discrimination among individual DNA hairpin molecules at single-nucleotide resolution using an ion channel. Structure of Staphylococcal alpha-hemolysin, a heptameric transmembrane pore. Characterization of individual polynucleotide molecules using a membrane channel. Detecting SNPs using a synthetic nanopore. Single-molecule analysis of DNA–protein complexes using nanopores. Protein biosensors based on biofunctionalized conical gold nanotubes. Characterization of nucleic acids by nanopore analysis. Incubation of 10 units of Klenow with supercoiled plasmid DNA produced no nicked molecules after 20 hours at 37 ☌ as determined by agarose gel electrophoresis analysis.Deamer, D. The enzyme is greater than 98 % pure as indicated by SDS-polyacrylamide gel electrophoresis and contains no detected endonuclease activity.

Dissolve 0.1 - 4 μg of digested DNA in 1x Reaction Buffer supplemented with 40 μM each dNTPĥ00 mM Tris-HCl pH 7.6 at 25 ☌, 50 mM MgCl 2 and 10 mM DTT.Įxcessive amounts of enzyme or longer reaction times may result in recessed ends due to the 3'→5' exonuclease activity of the enzyme.

Klenow retains the polymerization fidelity of the holoenzyme without degrading 5' termini. The enzyme lacks the 5'→3' exonuclease activity of intact DNA polymerase I. Klenow Fragment is the large fragment of DNA Polymerase I that retains its 5'→3' polymerase, 3'→5' exonuclease and strand displacement activities.

Removal of 3' overhangs to form blunt ends.

Fill-in of 5' overhangs to form blunt ends.Unit Definition: One unit is defined as the amount of enzyme required to convert 10 nmoles of dNTPs to an acid insoluble form in 30 minutes at 37 ☌.įorm: liquid (Supplied in 100 mM KPO 4 pH 6.5, 1 mM DTT and 50 % glycerol) RNA Labeling Selector (Kits & Nucleotides).DNA Labeling Selector (Kits & Nucleotides).Privacy Policy for Jena Bioscience's Facebook Page.Poly (A) carrier RNA-based RNA purification.3'-End RNA Labeling (T4 RNA Ligase 1-based).Random RNA Labeling ( in vitro Transcription-based).Kits for nucleoside base-modified mRNA Synthesis.Click Chemistry-based RNA/cRNA Labeling.Click Chemistry-based DNA/cDNA Labeling.Quantifoil® Holey Carbon Films with 2 nm Continuous Carbon.Mercurated and Selenium-containing Nucleotides.JBS Tungsten Cluster Derivatization Kits.Cryo and Room Temperature Crystallography.in Posttranslational Modification Analysis Immobilized Nucleotides for Affinity Chromatography.Stable Isotope labeled Nucleosides & Nucleotides.on Proteins/Enzymes - Sulfonyl Fluoride Probes Cancer and Proliferation Marker Nucleosides.

Klenow fragment dna polymerase i