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)
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• 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,50% GlycerolpH 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 assaysBsu 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, 1115–1121.3582 3585