Whole Cell Project Database -Purification-

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

Lot Information:

Purification Lot: PC011150-41-01-05
sPlasmid ID: PC011150-41
Plasmid Vector: pET-11a
Label: Native
Person: Ohmori M.
Purified Date: 04/05/2005
Total Protein Yield: 15.1 mg
Final Volume: 1.39 mL
Conc.: 10.9 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

03/02/2005BL21(DE3)6 L
03/15/2005BL21(DE3)6 L

Cell Lysis and Heat Treatment:

41.8g of E. coli cells
Ultra Sonication with 20mM Tris-HCl + 50mM 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
Resource ISO 6ml50 mM NaPi (pH 7.0), 1.5 M (NH4)2SO450.78 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)40.08 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)40.12 M NaPi
HiLoad 16/60 Superdex 200 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.588 ml
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)6