Imperial College/17 August 2008
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=====Monday===== | =====Monday===== | ||
- | Prepare plates and Media for later in the week | + | *Prepare plates and Media for later in the week |
- | Transform ''E.coli'' Xl1-blue with C0012 (LacI) from the registry again | + | *Transform ''E.coli'' Xl1-blue with C0012 (LacI) from the registry again |
=====Tuesday===== | =====Tuesday===== | ||
- | Check LacI transformants | + | *Check LacI transformants |
- | Miniprep and digest 0.1.01 (Double Terminator) and 0.1.02 (Chloraphenicol Reisitance) | + | *Miniprep and digest 0.1.01 (Double Terminator) and 0.1.02 (Chloraphenicol Reisitance). |
+ | **Ligate together to form construct 0.1.14 and transform into Xl1-Blue ''E.coli'' | ||
=====Wednesday===== | =====Wednesday===== | ||
- | Check ''E.coli'' transformed with construct 0.1.14 for growth and by PCR (if growth was succesful) | + | *Check ''E.coli'' transformed with construct 0.1.14 for growth and by PCR (if growth was succesful) |
- | If primers have arrived, PCR clone LacI, XylR, Spectinomycin, EpsE modified integration sequnce and AmyE integration sequence into Biobricks | + | *If primers have arrived, PCR clone LacI, XylR, Spectinomycin, EpsE modified integration sequnce and AmyE integration sequence into Biobricks |
=====Thursday===== | =====Thursday===== | ||
- | If grown, mini/midi-prep a few colonies from each plate and digest a sample with ''XbaI'' and ''SpeI'' to | + | *If grown, mini/midi-prep a few colonies from each plate and digest a sample with ''XbaI'' and ''SpeI'' to determine which plasmids have the insert in the correct orientation. |
- | If primers arrive today PCR cloning should be carried out | + | *If primers arrive today PCR cloning should be carried out |
=====Friday===== | =====Friday===== | ||
- | + | *If primers arrived Thursday check plates for growth and carry out minipreps | |
+ | |||
+ | *PCR check the minipreps that could be digested by ''XbaI'' and ''SpeI'' |
Revision as of 16:55, 15 August 2008
17th August 2008Wet LabB.subtilisCloningMonday
Tuesday
Wednesday
Thursday
Friday
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