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Plasmid & Protein Purification Database -Purification-

Species Sulfolobus tokodaii
Locus tag STS178
Products conserved hypothetical protein
Plasmid ID 0293
Vector  
Purification Lot STS178_1
Purified Date 2005.6.14-7.21
Protein conc. (mg/ml)  
Total volume (ml)  
Total protein (mg)  

Culture Lot:

date host cell scale label weight of bacteira (g)
        16.5

Cell Lysis and Heat Treatment:

16.5 g of E. coli.
75 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8, duty 50/1min, 5 times
Heat 10 min, 75 deg C (max 73 deg C), and then on ice 5 min.
Centrifuge at 100,000 g, 60 min.
 
 
 
 

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 3 1.1M AS
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 0.2M NaCl 3  
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1  
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Comment:

 

2011.10.20

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