This enzyme is inactivated by incubation at 75℃ for 20min.
Precautions：
Due to the lack of 3'→5' exonuclease activity, Bsu DNA Polymerase, Large Fragment cannot remove 3' overhangs and is therefore not suitable for generating blunt ends.Bsu DNA Polymerase, Large Fragment still retains 50% DNA polymerase activity at 25℃, which is twice the activity of Klenow fragment (3'→5' exo-) at the same temperature.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.
Instructions for Use：
1. Annealing of Primer/Template hybridsMix equimolar of single-stranded Primer and DNA template to a recommended final concentration of 10µM each, incubate at 90℃ for 1min, and then anneal by gradient cooling to 25℃ to form Primer/Template hybrids. We recommend using 's Annealing Buffer for DNA Oligos (5X) for the annealing according to the instructions of the product. The annealed duplex can be used directly for subsequent experiments, or stored at -20℃ for future use.2. Set up the following reaction on ice.ComponentVolumeFinal ConcentrationNuclease-free Water15µl-Annealed Primer/Template (10µM each)1µl0.5µM eachBsu Reaction Buffer (10X)2µl1XdNTP Mix (2.5mM each)1µl125µMBsu DNA Polymerase, Large Fragment (5U/µl)1µl0.25U/µlTotal Volume20µl-Note: When multiple reactions are required, prepare a master mix including all reagents except the Annealed Primer/template and then dispense to different nuclease-free PCR tubes. Finally, add Annealed Primer/Template hybrids to each tube.3. Incubate at 37℃ for an appropriate time period. The extension rate of this product is expected to be similar to that of conventional DNA polymerases, which is about 1000bp/min. For specific use, it is necessary to optimize the incubation time.4. Incubate at 75℃ for 20min to stop the reaction.
