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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST2435
Products  
Plasmid ID 0274
Vector  
Purification Lot ST2435
Purified Date  
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        20.5

Cell Lysis and Heat Treatment:

20.5 g of E. coli.
60 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50, on ice, 1 min on/1 min off, 7 times.
Heat 20 min, 80 deg C, then on ice 20 min.
Centrifuge at 100,000 rcf, 120 min.
100 ml of supernatant.
 
 
 

Purification Procedure:

Step Column Buffer Gradient 1 Gradient 2 FlowRate (ml/min) Eluted
1 ToyoScreen PPG 600M 5ml 20mM Tris-HCl(pH8.0),5mM B-ME,1.05M AS 1.05M AS 0M AS 4 0.60M AS
2 BioAssist Q 10x10 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 1M NaCl 5 300mM NaCl
3 Bio-Scale CHT10I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 250mM KPi 5 150mM KPi
4 G200SW(1inchx60) 10mM KPi(pH7.0),5mM B-ME,150mM NaCl     4 31min
5            
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Comment:

 

2011.10.20

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