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Available from Riken BioResource Center.

Plasmid & Protein Purification Database -Purification-

Species  
Locus tag APE0317
Products  
Plasmid ID R4081
Vector  
Purification Lot APE0317_2
Purified Date 2005.12.2-12.12
Protein conc. (mg/ml) 7.78
Total volume (ml) 0.6
Total protein (mg) 7.78

Culture Lot:

date host cell scale label weight of bacteira (g)
        93

Cell Lysis and Heat Treatment:

93 g of E. coli.
↓
400 ml of 20mM Tris-HCl (pH8.0), 5mM B-ME, 150mM NaCl.
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Sonication, output 8, duty 50/2min, 5 times.
↓
Heat 15 min, 85 deg C.
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Centrifuge at 100,000 rcf for 50 min.
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140 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,0.95M AS 1.05M AS 0.475M AS 4 0.8M AS
2 Resource Q 6 ml 20mM Tris-HCl(pH8.0),5mM B-ME 0M NaCl 0.4M NaCl 4 0.23M NaCl
3 HiLoad 16/60 Superdex200 20mM Tris-HCl(pH8.0),5mM B-ME,150mM NaCl     1 66ml/120ml
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Comment:

Products were confirmed by MS.

2011.10.20

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