Imperial College/17 August 2008

From 2008.igem.org

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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) then ligate together to form construct 0.1.14 and transform into Xl1-Blue ''E.coli''
+
*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 determin which plasmids have the insert in the correct orientation.  
+
*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=====
-
Check plates from Thursday for growth and/or PCR check the minipreps that could be digested by ''XbaI'' and ''SpeI''.
+
*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

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17th August 2008

Wet Lab

B.subtilis

Cloning

Monday
  • Prepare plates and Media for later in the week
  • Transform E.coli Xl1-blue with C0012 (LacI) from the registry again
Tuesday
  • Check LacI transformants
  • 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
  • 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
Thursday
  • 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
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
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