IGEM:MIT/2006/Notebook/2006-6-13: Difference between revisions
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#Created minipreps of the BAMT and SAMT from BL21 (6/9/06) | #Created minipreps of the BAMT and SAMT from BL21 (6/9/06) | ||
#*Used the protocol outlined: http://openwetware.org/wiki/Miniprep/Qiagen_kit | #*Used the protocol outlined: http://openwetware.org/wiki/Miniprep/Qiagen_kit | ||
#*Spun total of 3.6 mls of each- [as not many cells in culture, added (to 3.6) | #*Spun total of 3.6 mls of each- [as not many cells in culture, added more (to 3.6) and spun again] | ||
==To do== | ==To do== |
Revision as of 08:34, 13 June 2006
06/13/06 Lab
- Made glycerols of BAMT and SAMT from BL21 (6/9/06)
- Made two copies (4 total) of each by adding 1ml of each to glycerol stock
- Created minipreps of the BAMT and SAMT from BL21 (6/9/06)
- Used the protocol outlined: http://openwetware.org/wiki/Miniprep/Qiagen_kit
- Spun total of 3.6 mls of each- [as not many cells in culture, added more (to 3.6) and spun again]
To do
- Save glycerols of all cultures
- Miniprep two colonies of each construct
- Submit for sequencing with T7 forward and reverse primers (need to find primers).
- Check for transformants.
- If there are transformants, start cultures of colonies from each strain and construct combination.
Notes
Sequencing
- pET28a(+) genbank file from Novagen <- Is this is the vector that the constructs came in?
- This vector has a T7 promoter primer binding site but no T3, SP6, M13 Forward or M13 Reverse primer binding site.
- Does anyone have a T7 terminator primer?