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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0527
Products  
Plasmid ID 0712
Vector  
Purification Lot ST0527_1
Purified Date 2006.7.31
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg) 2.7

Culture Lot:

date host cell scale label weight of bacteira (g)
        19.5

Cell Lysis and Heat Treatment:

19.5 g of E. coli.
110 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 4 min, 10 times.
Heat 15min, 75 deg C.
Centrifuge at 100,000 rcf for 60min.
 
 
 
 

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,1.65M AS 1.65M AS 0M AS 4 1.03M AS
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 600mM NaCl 4 360mM NaCl
3 Bio-Scale CHT5I 10mM KPi(pH7.0),5mM B-ME,500mM KCl 10mM KPi 300mM KPi 2  
4 Resource Iso 6mL 20mM Tris-HCl(pH8.0),5mM B-ME,1.5M AS 1.5M AS 0M AS 4  
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Comment:

 

2011.10.20

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