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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST1480
Products  
Plasmid ID 0502
Vector  
Purification Lot ST1480_1
Purified Date 2004.8.2-
Protein conc. (mg/ml) 5.55
Total volume (ml) 0.4
Total protein (mg) 2.22

Culture Lot:

date host cell scale label weight of bacteira (g)
        30.5

Cell Lysis and Heat Treatment:

30.5 g of E. coli.
130 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 1 min, 7 times.
Heat 10min, 70 deg C.
Centrifuge at 100,000 rcf for 90min.
120 ml of supernatant.
 
 
 

Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Phe 6ml 20mM Tris-HCl(pH8.0),5mM B-ME,1.2M AS 1.2M AS 0M AS 6 0.22M AS
2 Bio-Scale CHT2I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 0.7 166.8mM KPi
3   20mM Tris-HCl(pH8.0),5mM B-ME        
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by MS.

2011.10.20

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