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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0119
Products  
Plasmid ID 4407
Vector  
Purification Lot APE0119
Purified Date 2005.9.29-10.13
Protein conc. (mg/ml) 12.2
Total volume (ml) 1.15
Total protein (mg) 14.3

Culture Lot:

date host cell scale label weight of bacteira (g)
        43.8

Cell Lysis and Heat Treatment:

43.8 g of E. coli.
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200 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
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Sonication output 8, duty 50/2min, 5 times.
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Heat 15 min 90 deg C.
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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,1.2M AS 1.2M AS 0M AS 5 0.8M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.8M NaCl 5 0.47M NaCl
3 Bio-Scale CHT5I 10mM KPi(pH7.0),5mM B-ME 10mM Kpi 300mM Kpi 0.9 174mM Kpi
4 HiLoad 16/60 Superdex 75       1 48ml/120ml
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Comment:

 

2011.10.20

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