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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST0944
Products hypothetical proliferating cell nuclear antigen
Plasmid ID 720
Vector  
Purification Lot ST0944
Purified Date 2006.11.28
Protein conc. (mg/ml) 36.77
Total volume (ml) 1.50
Total protein (mg) 55.1

Culture Lot:

date host cell scale label weight of bacteira (g)
        12.6

Cell Lysis and Heat Treatment:

12.6 g of E. coli.
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Sonication, output 8, duty 50/3 min, 5 times.
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Heat 10 min, 75 deg C.
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Centrifuge at 100,000 rcf for 60 min.
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Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Iso 6mL 20mM Tris-HCl(pH8.0),5mM B-ME,1.8M AS 1.8M AS 0M AS 6 1.53M AS
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 1.5M NaCl 4 310mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 86min
4            
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6            
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9            

Comment:

Products in the main band was confirmed by Trypsin-PMF.

2011.10.20

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