Whole Cell Project Database -Purification-

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

Lot Information:

Purification Lot: PC011150-41-01-03
sPlasmid ID: PC011150-41
Plasmid Vector: pET-11a
Label: Native
Person: Ebihara A.
Purified Date: 05/27/2003
Total Protein Yield: 16.2 mg
Final Volume: 0.7 mL
Conc.: 23.1 mg/mL

Final SDS-PAGE:

Culture Lot:

date

host cell

scale

SDS-PAGE

10/07/2001BL21(DE3)16 L

Cell Lysis and Heat Treatment:

39.4g 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)2SO45flowthrough and 0.9 M AS
Resource ISO 6ml50 mM NaPi (pH 7.0), 1.35 M (NH4)2SO451.0 M AS
Resource Q 6ml20 mM Tris-HCl (pH 8.0)50.08 M NaCl
Hydroxyapatite CHT10-I10 mM NaPi (pH 7.0)4100 mM NaPi
Mono Q 1ml20 mM Tris-HCl (pH 8.0)10.1 M NaCl
HiLoad 16/60 Superdex 75 pg20 mM Tris-HCl (pH 8.0), 150 mM NaCl0.5117 min (Mw59 K)
HiPrep 26/10 Desalting 53ml20 mM Tris-HCl (pH 8.0)7