Melbourne/Secondary Reagent TAE
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[[Melbourne/Protocols for Secondary Reagents|<return to list of secondary methods>]] [[Melbourne| <team home page>]] | [[Melbourne/Protocols for Secondary Reagents|<return to list of secondary methods>]] [[Melbourne| <team home page>]] | ||
===Preparation of TAE Buffer=== | ===Preparation of TAE Buffer=== | ||
| + | |||
===Uses=== | ===Uses=== | ||
| + | *Buffer for gel electrophoresis | ||
| + | |||
===Ingedients:=== | ===Ingedients:=== | ||
| - | * | + | Per Liter: |
| + | |||
| + | *242g Tris [[Melbourne/primary Tris|TRIS]] | ||
| + | *57.1ml glacial acetic acid | ||
| + | *100ml 0.5 M [[Melbourne/primary EDTA|EDTA]] (pH 8.0) | ||
| + | *Sterile water to 1L | ||
| + | |||
===Method:=== | ===Method:=== | ||
| - | # | + | #Make concentrated stock solution by mixing ingredients using a magnetic stirrer. |
| + | #Make to 1X from concentrated stock solution by adding sterile water. | ||
| + | |||
| + | Working Solution: | ||
| + | |||
| + | *0.04M Tris-acetate | ||
| + | *0.001M [[Melbourne/primary EDTA|EDTA]] | ||
| + | |||
| + | ===Note:=== | ||
| + | TAE has low buffering capacity and becomes exhausted during extended electrophoresis. Recirculation of buffer may be necessary. TBE buffer gives equally good resolution of DNA fragments but has significantly higher buffering capacity, making recirculation of buffer unnecessary. | ||
| + | |||
| + | ===References:=== | ||
| + | *T. Maniatis, E. G. Fritsch, J. Sambrook. Molecular Cloning, a Laboratory Manual. | ||
Revision as of 08:33, 29 September 2007
<return to list of secondary methods> <team home page>
Contents |
Preparation of TAE Buffer
Uses
- Buffer for gel electrophoresis
Ingedients:
Per Liter:
Method:
- Make concentrated stock solution by mixing ingredients using a magnetic stirrer.
- Make to 1X from concentrated stock solution by adding sterile water.
Working Solution:
- 0.04M Tris-acetate
- 0.001M EDTA
Note:
TAE has low buffering capacity and becomes exhausted during extended electrophoresis. Recirculation of buffer may be necessary. TBE buffer gives equally good resolution of DNA fragments but has significantly higher buffering capacity, making recirculation of buffer unnecessary.
References:
- T. Maniatis, E. G. Fritsch, J. Sambrook. Molecular Cloning, a Laboratory Manual.
