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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0457
Products  
Plasmid ID 4018
Vector  
Purification Lot APE0457_4
Purified Date 2005.6.6-6.16
Protein conc. (mg/ml) 3.63
Total volume (ml) 0.74
Total protein (mg) 2.69

Culture Lot:

date host cell scale label weight of bacteira (g)
        41.7

Cell Lysis and Heat Treatment:

41.7 g of E. coli.
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Sonication, output 8, duty 50/2min, 7 times.
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Heat 10 min, 85-90 deg C. It takes 10 min to 85 deg C.
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Centrifuge at 100,000 rcf for 50 min.
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190 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 6 1.27M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.5M NaCl 6 0.37M NaCl
3 Bio-Scale CHT2I 10mM KPi(pH7.0),5mM B-ME 10mM KPi 250mM KPi 1 173.8mM KPi
4 Superdex200HR10/30 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     0.5 26min
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Comment:

 

2011.10.20

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