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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST2362
Products  
Plasmid ID 0587
Vector  
Purification Lot ST2362
Purified Date 2006.4.3-
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        23.1

Cell Lysis and Heat Treatment:

23.1 g of E. coli.
110 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/3 min, 10 times.
Heat 15 min, 75 deg C.
Centrifuge at 100,000 rcf, 50 min.
92 ml of supernatant.
 
 
 

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,2M AS 2M AS 0M AS 5 1.71M AS
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 2M NaCl 4  
3            
4            
5            
6            
7            
8            
9            

Comment:

A contamination of 200 kDa was removed. Because of little of gain, procedure was abandoned.

2011.10.20

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