IGEM:MIT/2006/Notebook/2006-6-13: Difference between revisions
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#*Added 250µl of P2 and inverted a couple of times | #*Added 250µl of P2 and inverted a couple of times | ||
#*Added 350µl of N3 and centrifuged for 10 min | #*Added 350µl of N3 and centrifuged for 10 min | ||
#Created additional liquid cultures of colonies from Top10(TK) plates | |||
#*3 from each of SAMT, BAMT, and BSMT from 6/12/06. | |||
==To do== | ==To do== |
Revision as of 09:00, 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 (let sit for 30 min)
- 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]
- Added 250µl of P1 and vortexed
- Added 250µl of P2 and inverted a couple of times
- Added 350µl of N3 and centrifuged for 10 min
- Created additional liquid cultures of colonies from Top10(TK) plates
- 3 from each of SAMT, BAMT, and BSMT from 6/12/06.
To do
- 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?