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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST1970
Products  
Plasmid ID R0249
Vector  
Purification Lot ST1970_4
Purified Date 2005.6.9-6-16
Protein conc. (mg/ml) 14.73
Total volume (ml) 1.02
Total protein (mg) 15.03

Culture Lot:

date host cell scale label weight of bacteira (g)
        20.3

Cell Lysis and Heat Treatment:

20.3 g of E. coli.
↓
100 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
↓
Sonication, output 8, duty 50/2 min, 5 times.
↓
Heat 30 min, 80 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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93 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,1.35M AS 1.35M AS 0M AS 4 Flow through
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.5M NaCl 5 0.07M NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.4),5mM B-ME,150mM NaCl     1 65ml
4            
5            
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9            

Comment:

Products were confirmed by MS.

2011.10.20

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