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Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag ST0387
Products hypothetical proliferating cell nuclear antigen
Plasmid ID 0321
Vector  
Purification Lot ST0387
Purified Date  
Protein conc. (mg/ml) 3.90
Total volume (ml) 0.20
Total protein (mg) 0.78

Culture Lot:

date host cell scale label weight of bacteira (g)
        6.92

Cell Lysis and Heat Treatment:

6.92 g of E. coli.
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60 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/1min, 5 times.
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Heat 10 min 70 deg C and then on ice for 10min.
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Centrifuge at 100,000 rcf for 2 hours at 4 deg C.
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126 ml of supernatant.
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Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Bio-Scale CHT2I 2mL 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 1 100mM KPi
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 1M NaCl 6 300mM NaCl
3 Mono Q 1mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 500mM NaCl 1 360mM NaCl
4 Superdex 200 HR 10/30 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     0.5  
5 Resource Phe 6mL 20mM Tris-HCl(pH8.0),5mM B-ME,1.05M AS 1.05M AS 0M AS 2 700mM AS
6 Superdex 200 HR 10/30 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     0.5  
7            
8            
9            

Comment:

 

2011.10.20

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