Whole Cell Project Database -Purification-

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Thermus thermophilus HB8  TTHA1150
DNA polymerase beta family (X family)

Lot Information:

Purification Lot: PC011150-41-01-06
sPlasmid ID: PC011150-41
Plasmid Vector: pET-11a
Label: Native
Person: Manzoku M.
Purified Date: 08/10/2005
Total Protein Yield: 8.1 mg
Final Volume: 0.84 mL
Conc.: 9.6 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

06/21/2005BL21(DE3)9 L

Cell Lysis and Heat Treatment:

27.6g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 500mM NaCl (pH8.0)
Heat Treatment at 70°C for 10 min
Ultra Cengrifuge at 200,000G for 1 hr

Purification Procedure:

ColumnBufferFlow Rate (mL/min)Eluted
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10 
Super Q Toyopearl 650M (PACKED)20 mM Tris-HCl (pH 8.0)860mM NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)485mM NaCl
Hydroxyapatite CHT10-I10 mM KPi (pH 7.0)3160mM KPi
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl0.576.4 mL