Whole Cell Project Database -Purification-

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

Lot Information:

Purification Lot: PC011150-41-02-04
sPlasmid ID: PC011150-41
Plasmid Vector: pET-11a
Label: SeMet
Person: Ohmori M.
Purified Date: 10/22/2004
Total Protein Yield: 4.5 mg
Final Volume: 0.612 mL
Conc.: 7.3 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

09/01/2004B834(DE3)pLysS12 L

Cell Lysis and Heat Treatment:

36.5g 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 30 ml20 mM Tris-HCl (pH 8.0)40.08 M NaCl
Resource Q 6ml20 mM Tris-HCl (pH 8.0)40.10 M NaCl
Hydroxyapatite CHT10-I10 mM KPi (pH 7.0)30.13 M KPi
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 200 mM NaCl0.579 ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)10