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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST2030
Products  
Plasmid ID R0373
Vector  
Purification Lot ST2030
Purified Date 2006.5.15-5.25
Protein conc. (mg/ml) 18.30
Total volume (ml) 1.43
Total protein (mg) 26.17

Culture Lot:

date host cell scale label weight of bacteira (g)
        40.0

Cell Lysis and Heat Treatment:

40.0 g of E. coli.
↓
210 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
↓
Sonication, output 8, duty 50/2 min, 5 times.
↓
Heat 15 min, 75 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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180 ml of supernatant.
↓
 
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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,2.0M AS 2.0M AS 0M AS 5 1.9 M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.6M NaCl 5 0.09M NaCl
3 HiLoad 16/60 Superdex200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 75mL/120mL
4            
5            
6            
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8            
9            

Comment:

Products was confirmed by Trp-MS.

2011.10.20

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