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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0546
Products  
Plasmid ID 4086
Vector  
Purification Lot APE0546
Purified Date  
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:

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

Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1            
2            
3            
4            
5            
6            
7            
8            
9            

Comment:

While target products were observed in HP step, there was no protein in S step. Further purification was abandoned.

2011.10.20

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