IGEM:MIT/2006/Notebook/2006-7-12: Difference between revisions
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Ran a gel of the osmY promoter (both truncated and full regions) with controls. We did not see any bands. | Ran a gel of the osmY promoter (both truncated and full regions) with controls. We did not see any bands. | ||
==Email requests progress== |
Revision as of 12:22, 12 July 2006
To do
- Check biobrick sequence results against GenBank sequences
- Make liquid cultures from YYC912 plates
- Continue GC standards/readings work
- Miniprep and digest/sequence (both) mutagenesis transformants
- run a gel of and then biobrick the stationary phase promoter PCR product
- Hook up each biobrick coding region (SAMT, BAMT, BSMT, ATF1) to a Rbs, Promotor, and Terminator
Biobrick Progress
Made a miniprep from the liquid culture made up last night carrying the terminator B0015. Also made a glycerol of the B0015 culture.
Did the following digest for later ligation between the SAMT coding region and B0015 and between the BSMT coding region and B0015 (the other coding regions are in the middle of testing after site-directed mutagenesis).
SAMT
30.2 uL H20
5 uL Buffer 2 (vortexed quickly)
.5 uL BSA
13.25 uL 60.4 ng/uL DNA
.5 uL EcoRI
.5 uL SpeI
BSMT
32.25 uL H20
5 uL Buffer 2 (vortexed quickly)
.5 uL BSA
10 uL 71.2 ng/uL DNA (ran out, ideal would have been 11.25 uL)
.5 uL EcoRI
.5 uL SpeI
B0015
39.5 uL H20
5 uL Buffer 2 (vortexed quickly)
.5 uL BSA
4 uL 203.9 ng/uL DNA
.5 uL EcoRI
.5 uL XbaI
- Note: We are looking for promoters already attached to RBSs, so we can skip that assembly process.
Prep for GC
Grew up liquid cultures of Dudareva's BSMT and SAMT in LB Kan as well as BL21 in LB as a control
Mutagenesis Progress
There were no ATF1 colonies. There were 2 BAMT colonies. We made two liquid cultures from the 2 BAMT colonies.
osmY promoter
Ran a gel of the osmY promoter (both truncated and full regions) with controls. We did not see any bands.