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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0397
Products  
Plasmid ID 0020-3
Vector  
Purification Lot ST0397_3
Purified Date 2005.7.11-7.21
Protein conc. (mg/ml) 13.09
Total volume (ml) 0.86
Total protein (mg) 11.26

Culture Lot:

date host cell scale label weight of bacteira (g)
        10.5

Cell Lysis and Heat Treatment:

10.5 g of E. coli.
↓
70ml Buffer/ B-ME 5ul
↓
Sonication, output 8/duty 50, 2 min, 5 times.
↓
Heat 15min, 75 deg C.
↓
Centrifuge at 100,000 rcf for 60min.
↓
 
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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,1.50M AS 1.5M AS 0M AS 4 1.4M AS
2 BioAssist Q 10X10 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 200mM NaCl 2 120mM NaCl
3 HiLoad 16/60 Superdex 75 20mM Tris-HCl(pH8.0),5mM B-ME     1 67ml/120ml
4            
5            
6            
7            
8            
9            

Comment:

Products were confirmed by MS in entire sequence.

2011.10.20

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