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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0385
Products  
Plasmid ID 0019
Vector  
Purification Lot ST0385_1a
Purified Date  
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        14.2

Cell Lysis and Heat Treatment:

14.2 g of E. coli.
25 ml of buffer.
Sonication, output 8, duty 50/1min, 5 times, on ice.
Heat 20 min 80 deg C.
Heat 20 min 80 deg C heat buffer 35ml, and then on ice.
Centrifuge at 100,000 rcf for 2 hours.
38.5 ml of supernatant.
 
 

Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 Resource Iso 6mL   1.5M AS 0M AS 5 M AS
2 BioAssist Q 10X10   0M NaCl 500mM NaCl 5  
3 Superdex 200 HR1 0/30          
4 Bio-Scale CHT10I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 250mM KPi 4  
5            
6            
7            
8            
9            

Comment:

At the end of purification 2, solution was divided into three fractions, a, b, and c, and the purified individually.

2011.10.20

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