Bsu DNA Polymerase I, Large Fragment is a product of the Bacillus subtilis DNA polymerase I which lacks the N-terminal exonuclease domain (1-296 amino acids). It retains the 5´→ 3´ polymerase activity of DNA polymerase I but lacks the 5´→ 3´ exonuclease activity. This large fragment also lacks 3´→ 5´ exonuclease activity (1).
Note:Glycerol acts as a cryoprotectant and protein stabilizer when added to the storage buffer of proteins and enzymes. However, in certain circumstances, it is preferred to omit glycerol from the buffer. This includes instances where the presence of glycerol may interfere, such as lyophilization, high-throughput instruments with sensitive fluidics or primary cell cultures.For our glycerol free products, it is recommended to use immediately upon thaw as without glycerol there will be significant activity loss with freeze/thaw cycles.
Applications• Strand displacement DNA synthesis (2)• Random primer labeling• Second strand cDNA synthesis• dA-tailing
Protein PurityThe physical purity of Bsu DNA Polymerase I, Large Fragment is ≥99% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).
Product SourceE. coli strain expressing Bsu Polymerase I gene lacking the N-terminal exonuclease domain.
Product Includes• Glycerol Free Bsu DNA Polymerase I, Large fragment• 10x Bsu DNA Polymerase I reaction buffer
1x Bsu Polymerase I reaction buffer10 mM Tris-HCl50 mM KCl10 mM MgCl21 mM DTTpH 7.9 @ 25°C
Storage Buffer50 mM Tris-HCl50 mM KCl1 mM DTT0.1 mM EDTA,pH 7.5 @ 25ºC
Storage Temperature–20ºC
Heat inactivation70°C for 20 min
Unit DefinitionOne unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTP into acid-insoluble form in 30 minutes at 37º C.
Quality Control assaysGlycerol Free Bsu Polymerase I, Large fragment is free from detectable nuclease activities. Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product.
References1. Okazaki, T. et al. (1964) J. Biol. Chem. 239, 259–268.2. Piepenburg, O. et al. (2006) PLOS Biology, 4, 1115–1121.3582G 3585G
