pfu polymerase, Promega or Stratagen

pyrobest from Takara

T7 endonuclease, New England Biolabs

T4 DNA ligase

Biokeystone Co

US patent 5,549,806, 5904826, http://www.6mgel.com/febe.htm

Invitrogen

E-gel:

5,865,974 and 5,582,702

中国工业微生物菌种保藏管理中心: http://www.china-cicc.org/

中国科学院微生物研究所(AS)

中国科学院武汉病毒研究所(AS-IV)

中国农业科学院土壤与肥料研究所(ISF)

中国食品发酵工业研究所(IFFI)

中国医学科学院皮肤病研究所(ID)，南京

卫生部药品生物制品检定所(NICPBP)

中国预防医学科学院病毒研究所

中国医学科学院医药生物技术研究所

四川抗生素研究所(SIA)，四川

华北制药厂抗生素研究所(IANP), 石家庄

农业部兽药监察研究所(NCIVBP)

中国林业科学院林业研究所(RIF)

http://www.alphabiosciences.com (in Baltimore).

You can wash your resin with 0.5N NaOH 30min for rapid regeneration. And use the same resin for the same protein purification…

For information :

Handling :

Ni-NTA matrices are stable under a wide variety of conditions and need not be refrigerated,
except to inhibit growth of microorganisms for long-term storage. After use they should be
washed for 30 minutes with 0.5M NaOH. Ni-NTA matrices should be stored in
30% ethanol to inhibit microbial growth. The matrix can be stored for up to one week in
any of the denaturing buffers.

Reuse of Ni-NTA Resin

The reuse of Ni-NTA resin depends on the nature of the sample and should only be performed
with identical recombinant proteins. Based on the experience of Hoffmann-La Roche Ltd.
(Basel, Switzerland), who have purified more than 100 different proteins on Ni-NTA resin,
we recommend a maximum of 5 runs per column.

If the Ni-NTA Agarose changes from light blue to brownish-gray, the following regeneration
procedure is recommended.
Procedure:
1. Wash the column with 2 volumes of Regeneration Buffer (6 M GuHCl, 0.2 M acetic
acid).
2. Wash the column with 5 volumes of H2O.
3. Wash the column with 3 volumes of 2% SDS.
4. Wash the column with 1 volume of 25% EtOH.
5. Wash the column with 1 volume of 50% EtOH.
6. Wash the column with 1 volume of 75% EtOH.
7. Wash the column with 5 volumes of 100% EtOH.
8. Wash the column with 1 volume of 75% EtOH.
9. Wash the column with 1 volume of 50% EtOH.
10. Wash the column with 1 volume of 25% EtOH.
11. Wash the column with 1 volume of H2O.
12. Wash the column with 5 volumes of 100 mM EDTA, pH 8.0.
13. Wash the column with H2O.
14. Recharge the column with 2 volumes of 100 mM NiSO4.
15. Wash the column with 2 volumes of H2O.
16. Wash the column with 2 volumes of Regeneration Buffer.
17. Equilibrate with 2 volumes of a suitable buffer (e.g., Buffer A