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Plasmid & Protein Purification Database -Purification-

Species Aeropyrum pernix
Locus tag APE0012
Products ribokinase
Plasmid ID 4375
Vector  
Purification Lot APE0012_1
Purified Date 2005.2.8-
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        50.0

Cell Lysis and Heat Treatment:

50.0 g of E. coli.
↓
200 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
↓
Sonication, output 8, duty 50/2 min, 6 times.
↓
Heat 15 min, 75 deg C.
↓
Centrifuge at 100,000 rcf, 60 min.
↓
160 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 0.32M AS
2            
3            
4            
5            
6            
7            
8            
9            

Comment:

 

2011.10.20

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