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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST1432
Products  
Plasmid ID R0223
Vector  
Purification Lot ST1432
Purified Date  
Protein conc. (mg/ml) 15.95
Total volume (ml) 0.92
Total protein (mg) 14.67

Culture Lot:

date host cell scale label weight of bacteira (g)
        27.2

Cell Lysis and Heat Treatment:

27.2 g of E. coli.
140 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
Sonication, output 8/duty 50, 5 min, 2 times.
Heat 15min, 75 deg C.
Centrifuge at 100,000 rcf for 60min.
120 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 1.5M AS
2 Resource S 6 ml 50mM MES(pH6.0),5mM B-ME 0M NaCl 0.4M NaCl 4 0.11M NaCl
3 HiLoad 16/60 Superdex200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 82ml/120ml
4            
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Comment:

Products were confirmed by Trp-Mass.

2011.10.20

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