Reagents required for de novo gene synthesis
pfu polymerase, Promega or Stratagen
pyrobest from Takara
T7 endonuclease, New England Biolabs
T4 DNA ligase
Tags: Biotechnologypfu polymerase, Promega or Stratagen
pyrobest from Takara
T7 endonuclease, New England Biolabs
T4 DNA ligase
Tags: BiotechnologyBiokeystone Co
US patent 5,549,806, 5904826, http://www.6mgel.com/febe.htm
Invitrogen
E-gel:
5,865,974 and 5,582,702
Tags: Biotechnology中国工业微生物菌种保藏管理中心: http://www.china-cicc.org/
| 中国普通微生物菌种保藏中心CGMCC(归口中国科学院) |
| 中国农业微生物菌种保藏中心ACCC(归口中国农业科学院) |
| 中国工业微生物菌种保藏中心CICC(归口轻工业总会) |
| 中国医学微生物菌种保藏中心CMCC(归口卫生部) |
| 中国抗生素微生物菌种保藏中心CACC(国家医药管理局) |
| 中国兽医微生物菌种保藏中心CVCC(归口农业部) |
| 中国林业微生物菌种保藏中心CFCC(归口中国林业科学院) |
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
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