IGEM:MIT/2006/Notebook/2006-6-13: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
| Line 7: | Line 7: | ||
#*Added 250µl of P1 and vortexed | #*Added 250µl of P1 and vortexed | ||
#*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 | #*Added 350µl of N3 and centrifuged for 10 min | ||
==To do== | ==To do== | ||
Revision as of 08:54, 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]
- 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
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?
