IGEM:MIT/2006/Notebook/2006-6-13

06/13/06 Lab

1. 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
2. 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 as many cells in culture, added more and spun again.

To do

1. Save glycerols of all cultures
2. Miniprep two colonies of each construct
3. Submit for sequencing with T7 forward and reverse primers (need to find primers).
4. Check for transformants.
5. 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?