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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag ST0151
Products  
Plasmid ID 0401
Vector  
Purification Lot ST0151
Purified Date 2007.2.14
Protein conc. (mg/ml) 1.11
Total volume (ml) 11.0
Total protein (mg) 12.2

Culture Lot:

date host cell scale label weight of bacteira (g)
        24.3

Cell Lysis and Heat Treatment:

24.3 g of E. coli cell.
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140 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 50mM NaCl.
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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,1.05M AS 1.05M AS 0M AS 4 50mM AS
2 Resource Q 6mL 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 400mM NaCl 4 80mM NaCl
3 HiLoad 16/60 Superdex 200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 75mL/120mL
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Comment:

Solusion became clouded after application of an ion exchange column. Supernatant of centrifuged solusion was used for further step. Products were confirmed by Trp-Mass.

2011.10.20

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