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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0408
Products  
Plasmid ID 411
Vector  
Purification Lot ST0408_2
Purified Date 2006.12.18
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        43.8

Cell Lysis and Heat Treatment:

43.8 g of E. coli.
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Sonication, output 8/duty 50, 3 min, 5 times.
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Heat 10min, 70 deg C.
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Centrifuge at 100,000 rcf for 60min.
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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,1.50M AS 1.50M AS 0M AS 6 870mM AS
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 600mM NaCl 4 260mM NaCl
3 Bio-Scale CHT5I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 500mM KPi 2 85.7mM KPi
4 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 74ml
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Comment:

 

2011.10.20

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