DNA Polymerase Alpha, Calf Thymus

Description

• Multi-subunit enzyme complex isolated from calf thymus glands by immunoaffinity chromatography (1)
• Exhibits DNA polymerase and primase activities
• For use in DNA replication experiments requireing a eukaryotic source of DNA polymerase alpha-primase

Unit Definition

One unit is the amount of enzyme required to incorporate 1nmol of total nucleotide into acid-insoluble form at 37°C in 60 minutes

Storage Buffer

20 mM Tris-HCl (pH 8.5)
50 mM KCl
0.25 mM EDTA
1 mM dithiothreitol
50% (v/v) glycerol

Assay Conditions

60 mM Tris-HCl (pH 8.0)
5 mM magnesium acetate
0.3 mg/ml bovine serum albumin
0.1 mM each of dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [³H]dTTP
10 µg activated calf thymus DNA
Final volume of 50 µl

Quality Control

All preparations are assayed for contaminating endonuclease and exonuclease activities. The polymerase is also tested in primase-dependent DNA replicantion system (2)

Storage Conditions

Store at -20°C
Product shipped on dry ice