Whole Cell Project Database -Purification-

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

Lot Information:

Purification Lot: PC011150-41-01-02
sPlasmid ID: PC011150-41
Plasmid Vector: pET-11a
Label: Native
Person: Ebihara A.
Purified Date: 09/03/2001
Total Protein Yield: 5.5 mg
Final Volume: .37 mL
Conc.: 14.8 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

07/04/2001BL21(DE3)8 L

Cell Lysis and Heat Treatment:

23.37g 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), 1.35 M (NH4)2SO45.0Flowthrough and 0.89 M AS
Resource ISO 6ml50 mM NaPi (pH 7.0), 1.35 M (NH4)2SO45.00.89 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)5.00.072 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)5.0110 mM NaPi
Mono Q 1ml20 mM Tris-HCl (pH 8.0)1.00.10 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5 
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)7.0