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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0550
Products  
Plasmid ID R4333
Vector  
Purification Lot APE0550_5
Purified Date 2006.5.23-
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
         

Cell Lysis and Heat Treatment:

13.3 g of E. coli.
↓
73.5 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
↓
Sonication, output 8, duty 50/4 min, 8 times.
↓
Heat 10 min, 80 deg C.
↓
Centrifuge at 100,000 rcf for 60 min.
↓
 
↓
 
↓
 
 

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,1.5M AS 1.8M AS 0M AS 4 1.45M AS
2            
3            
4            
5            
6            
7            
8            
9            

Comment:

Cultured in a Se-Met containing medium.

2011.10.20

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