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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0365
Products  
Plasmid ID 0018
Vector  
Purification Lot ST0365
Purified Date 2007.2.14
Protein conc. (mg/ml) 37.4
Total volume (ml) 1.15
Total protein (mg) 43.0

Culture Lot:

date host cell scale label weight of bacteira (g)
        40.6

Cell Lysis and Heat Treatment:

40.6 g of E. coli.
210 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/5 min, 5 times.
Heat 20 min 75 deg C.
Centrifuge at 100,000 rcf for 45 min.
174 ml of supernatant.
 
 
 

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 500mM AS 5 1.63M AS
2 Resource S 6mL 50mM MES(pH6.0),5mM B-ME 0M NaCl 300mM NaCl 4 90mM NaCl
3 Bio-Scale CHT5I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 375mM KPi 2 189mM KPi
4 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 72ml/120ml
5            
6            
7            
8            
9            

Comment:

Products were confirmed by Trp-MS.

2011.10.20

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