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Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0319
Products  
Plasmid ID R4703
Vector  
Purification Lot APE0319_b
Purified Date  
Protein conc. (mg/ml) 14.14
Total volume (ml) 0.80
Total protein (mg) 11.3

Culture Lot:

date host cell scale label weight of bacteira (g)
        28.3

Cell Lysis and Heat Treatment:

28.3 g of E. coli.
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Sonication, output 8, duty 50/4min, 5 times.
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Heat 10 min, 80 deg C.
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Centrifuge at 100,000 rcf for 60 min.
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150 ml of supernatant.
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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,1.5M AS 1.5M AS 0M AS 3 1.17M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 2M NaCl 4 0.15M NaCl
3 HiLoad 16/60 Superdex200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 68min
4            
5            
6            
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8            
9            

Comment:

Maltimeric products. Because two peaks were obtaied at the step of Reso Q, products were gel purified independent two (a and b) ways. Products were eluted at 80 min in a procedure and at 68 min in b procedure. Products were able to purified without precipitation in the solution. Products of high molecular weight were also confirmed.

2011.10.20

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