User:University of Washington/4 August 2008
From 2008.igem.org
LuxR from pLac
-R0010 and E0240 ligation was transformed into DH5a strain.
-Original R0010 and E0240 ligation (from two? weeks ago) that had an inconclusive sequencing was sent in for sequencing again.
-Part I0462 (receieved from igem hq) was sent in for sequencing.
MG1655Z1
- MG1655Z1 from section 1 and 5 were miniprepped. Glycerol stock were made for both.
LuxR from AraC and TetR
- Ran gel for P1010 (AC x 3, Kan x 3) to combine the DNA into one tube.
- Restriction digest the P1010 only one on pSB1AC3, and the PCR Elowitz's promoter D29 with XbaI and PstI.
- mixed two reaction
dH2O(ul) | NEB3(ul) | BSA(ul) | DNA(ul) | XbaI(ul) | PstI(ul) | |
---|---|---|---|---|---|---|
P1010 (AC) | 32.5 | 5 | 0.5 | 10 | 1 | 1 |
D29 | 32.5 | 5 | 0.5 | 10 | 1 | 1 |
- incubated 37 degree Celcius about 1 hour
- 20 mins in 80 degree Celcius water bath
- stored in the fridge
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