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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0410
Products  
Plasmid ID 0412
Vector  
Purification Lot ST0410
Purified Date 2006.2.24
Protein conc. (mg/ml) 9.31
Total volume (ml) 1.25
Total protein (mg) 11.6

Culture Lot:

date host cell scale label weight of bacteira (g)
        8.0

Cell Lysis and Heat Treatment:

8.0 g of E. coli.
74.5 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 3 min, 10 times.
Heat 15min, 75 deg C.
Centrifuge at 100,000 rcf for 50min.
64 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,1.75M AS 1.75M AS 0M AS 4 1.49M AS
2 Resource S 6mL 50mM MES(pH6.0),5mM B-ME 0M NaCl 2M NaCl 4 120mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 65min
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Comment:

 

2011.10.20

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