KLENOW FRAGMENT MOLECULAR WEIGHT SERIAL
KLENOW FRAGMENT MOLECULAR WEIGHT FREE

Purity was assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the protein of interest band in the diluted sample. Klenow Fragment (3'→5' exo-) physical purity was evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver stain detection. Protein concentration of Klenow Fragment (3'→5' exo-) (OD 280) was determined by OD 280 absorbance. Reactions were incubated for 10 minutes at 37☌, plunged on ice, and analyzed using the method of Sambrook and Russell (Molecular Cloning, v3, 2001, pp.

Dilutions of enzyme were made in a glycerol (50%) containing Klenow (3'→5' exo-) storage solution and added to 50 μL reactions containing Calf Thymus DNA, 1X Blue Buffer, 3H-dTTP and 100 μM dNTPs.

KLENOW FRAGMENT MOLECULAR WEIGHT SERIAL

Klenow Fragment (3'→5' exo-) unit activity was measured using a 2-fold serial dilution method. Instructions for using Klenow Fragment (3'→5' exo-) are provided in the corresponding kit protocol in the resources below. The action of Klenow Fragment (3'→5' exo-) is also utilized in some fluorescent labeling reactions for microarray. This procedure is frequently used in preparing DNA libraries for next-generation sequencing. This process of dA-tailing (or dT-tailing) of the DNA is an important step in the process of ligating DNA adapters to DNA fragments. DNA polymerases lacking a 3'→5' exonuclease may add a single nontemplate-dependent nucleotide to the 3'-end of a sequence (tailing) with a preference for dATP depending on the reaction conditions and sequence context. This problem is overcome by introducing mutations that result in the Klenow Fragment (3'→5' exo-) enzyme that retains 5'→3' polymerase activity but lacks any exonuclease activity. coli DNA polymerase I can be undesirable, the 3'→5' exonuclease proofreading activity of Klenow Fragment can also be undesirable in some circumstances.

Just as the 5'→3' exonuclease activity of E. coli DNA polymerase I lacks this 5’-exonuclease activity but retains the 5'→3' polymerase activity and the 3'→5' exonuclease activity for removal of precoding nucleotides and proofreading.

KLENOW FRAGMENT MOLECULAR WEIGHT FREE

It is undesirable for DNA sequencing and other applications due to the ability of the 5’→3’ exonuclease to degrade flap structures, single-stranded and double-stranded DNA from free 53'→5'-ends. The 3'→5' nuclease activity is used during DNA repair and is necessary for enzyme applications such as nick translation. The main duty of the 5'→3' exonuclease activity is to remove the RNA primers at the 5' ends of newly synthesized DNA so that the polymerase activity can fill in the resulting gaps. coli DNA polymerase I makes it unsuitable for some applications. The holoenzyme possesses three activities: 5'→3' DNA-dependent DNA polymerase, 3'→5' proofreading and 3'→5' exonuclease (also known as flap endonuclease). DNA polymerase I is a single polypeptide chain consisting of ∼930 residues.

Klenow fragment molecular weight