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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST1969
Products  
Plasmid ID 248
Vector  
Purification Lot ST1969_2c
Purified Date 2005.2.23-2-28
Protein conc. (mg/ml) 9.97
Total volume (ml) 0.7
Total protein (mg) 6.98

Culture Lot:

date host cell scale label weight of bacteira (g)
        47.0

Cell Lysis and Heat Treatment:

47.0 g of E. coli.
↓
125 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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Sonication, output 8, duty 50/2 min, 7 times.
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Heat 10 min, 70 deg C.
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Centrifuge at 100,000 rcf, 60 min.
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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 6 0.97M AS
2 Resource S 6 ml 50mM MES(pH5.7),5mM B-ME 0M NaCl 2M NaCl 6 130mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.4),5mM B-ME,150mM NaCl     1 71min
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by MS.

2011.10.20

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