Imperial College/17 August 2008

From 2008.igem.org

July
MTWTFSS
  [http://2008.igem.org/wiki/index.php?title=Imperial_College/1_July_2008&action=edit 1] [http://2008.igem.org/wiki/index.php?title=Imperial_College/2_July_2008&action=edit 2] [http://2008.igem.org/wiki/index.php?title=Imperial_College/3_July_2008&action=edit 3] [http://2008.igem.org/wiki/index.php?title=Imperial_College/4_July_2008&action=edit 4] [http://2008.igem.org/wiki/index.php?title=Imperial_College/5_July_2008&action=edit 5] [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_July_2008&action=edit 6]
[http://2008.igem.org/Imperial_College/7_July_2008 7] [http://2008.igem.org/Imperial_College/8_July_2008 8] [http://2008.igem.org/Imperial_College/9_July_2008 9] [http://2008.igem.org/wiki/index.php?title=Imperial_College/10_July_2008&action=edit 10] [http://2008.igem.org/Imperial_College/11_July_2008 11] [http://2008.igem.org/wiki/index.php?title=Imperial_College/12_July_2008&action=edit 12] [http://2008.igem.org/wiki/index.php?title=Imperial_College/13_July_2008&action=edit 13]
[http://2008.igem.org/wiki/index.php?title=Imperial_College/14_July_2008&action=edit 14] [http://2008.igem.org/Imperial_College/15_July_2008 15] [http://2008.igem.org/Imperial_College/16_July_2008 16] [http://2008.igem.org/wiki/index.php?title=Imperial_College/17_July_2008&action=edit 17] [http://2008.igem.org/Imperial_College/18_July_2008 18] [http://2008.igem.org/wiki/index.php?title=Imperial_College/19_July_2008&action=edit 19] [http://2008.igem.org/wiki/index.php?title=Imperial_College/20_July_2008&action=edit 20]
[http://2008.igem.org/Imperial_College/21_July_2008 21] [http://2008.igem.org/Imperial_College/22_July_2008 22] [http://2008.igem.org/Imperial_College/23_July_2008 23] [http://2008.igem.org/Imperial_College/24_July_2008 24] [http://2008.igem.org/Imperial_College/25_July_2008 25] [http://2008.igem.org/wiki/index.php?title=Imperial_College/26_July_2008&action=edit 26] [http://2008.igem.org/wiki/index.php?title=Imperial_College/27_July_2008&action=edit 27]
[http://2008.igem.org/Imperial_College/28_July_2008 28] [http://2008.igem.org/Imperial_College/29_July_2008 29] [http://2008.igem.org/Imperial_College/30_July_2008 30] [http://2008.igem.org/wiki/index.php?title=Imperial_College/31_July_2008&action=edit 31]
August
MTWTFSS
        [http://2008.igem.org/Imperial_College/1_August_2008 1] [http://2008.igem.org/Imperial_College/2_August_2008 2] [http://2008.igem.org/wiki/index.php?title=Imperial_College/3_August_2008&action=edit 3]
[http://2008.igem.org/Imperial_College/4_August_2008 4] [http://2008.igem.org/Imperial_College/5_August_2008 5] [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_August_2008&action=edit 6] [http://2008.igem.org/Imperial_College/7_August_2008 7] [http://2008.igem.org/Imperial_College/8_August_2008 8] [http://2008.igem.org/wiki/index.php?title=Imperial_College/9_August_2008&action=edit 9] [http://2008.igem.org/Imperial_College/10_August_2008 10]
[http://2008.igem.org/Imperial_College/11_August_2008 11] [http://2008.igem.org/Imperial_College/12_August_2008 12] [http://2008.igem.org/Imperial_College/13_August_2008 13] [http://2008.igem.org/Imperial_College/14_August_2008 14] [http://2008.igem.org/Imperial_College/15_August_2008 15] [http://2008.igem.org/Imperial_College/16_August_2008 16] [http://2008.igem.org/Imperial_College/17_August_2008 17]
[http://2008.igem.org/Imperial_College/18_August_2008 18] [http://2008.igem.org/Imperial_College/19_August_2008 19] [http://2008.igem.org/Imperial_College/20_August_2008 20] [http://2008.igem.org/Imperial_College/21_August_2008 21] [http://2008.igem.org/Imperial_College/22_August_2008 22] [http://2008.igem.org/wiki/index.php?title=Imperial_College/23_August_2008&action=edit 23] [http://2008.igem.org/Imperial_College/24_August_2008 24]
[http://2008.igem.org/Imperial_College/25_August_2008 25] [http://2008.igem.org/Imperial_College/26_August_2008 26] [http://2008.igem.org/Imperial_College/27_August_2008 27] [http://2008.igem.org/Imperial_College/28_August_2008 28] [http://2008.igem.org/Imperial_College/29_August_2008 29] [http://2008.igem.org/wiki/index.php?title=Imperial_College/30_August_2008&action=edit 30] [http://2008.igem.org/Imperial_College/31_August_2008 31]
September
MTWTFSS
[http://2008.igem.org/Imperial_College/1_September_2008 1] [http://2008.igem.org/Imperial_College/2_September_2008 2] [http://2008.igem.org/Imperial_College/3_September_2008 3] [http://2008.igem.org/Imperial_College/4_September_2008 4] [http://2008.igem.org/Imperial_College/5_September_2008 5] [http://2008.igem.org/wiki/index.php?title=Imperial_College/6_September_2008&action=edit 6] [http://2008.igem.org/Imperial_College/7_September_2008 7]
[http://2008.igem.org/Imperial_College/8_September_2008 8] [http://2008.igem.org/Imperial_College/9_September_2008 9] [http://2008.igem.org/Imperial_College/10_September_2008 10] [http://2008.igem.org/Imperial_College/11_September_2008 11] [http://2008.igem.org/Imperial_College/12_September_2008 12] [http://2008.igem.org/wiki/index.php?title=Imperial_College/13_September_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=Imperial_College/14_September_2008&action=edit 14]
[http://2008.igem.org/Imperial_College/15_September_2008 15] [http://2008.igem.org/Imperial_College/16_September_2008 16] [http://2008.igem.org/Imperial_College/17_September_2008 17] [http://2008.igem.org/Imperial_College/18_September_2008 18] [http://2008.igem.org/Imperial_College/19_September_2008 19] [http://2008.igem.org/wiki/index.php?title=Imperial_College/20_September_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=Imperial_College/21_September_2008&action=edit 21]
[http://2008.igem.org/Imperial_College/22_September_2008 22] [http://2008.igem.org/Imperial_College/23_September_2008 23] [http://2008.igem.org/Imperial_College/24_September_2008 24] [http://2008.igem.org/Imperial_College/25_September_2008 25] [http://2008.igem.org/Imperial_College/26_September_2008 26] [http://2008.igem.org/wiki/index.php?title=Imperial_College/27_September_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=Imperial_College/28_September_2008&action=edit 28]
[http://2008.igem.org/Imperial_College/29_September_2008 29] [http://2008.igem.org/Imperial_College/30_September_2008 30]

17 August 2008

Wet Lab

B.subtilis

Monday
  • Prepare media and reagents for the protocol transformation 1 protocol
  • Prepare 2x10ml LB media in 100ml flasks
  • Prepare 2x100ml LB media in 1000ml flasks
  • Prepare 2x overnight cultures in the 10ml LB media flasks
  • Prepare a 1% agarose gel and run the digest prepared on friday
Tuesday
  • Prepare competent cells using the transformation 2 protocol, this time growing cells to an o.D.600
  • Prepare 1x20ml of LB media in a 200ml flask (autoclaved in the morning)
  • Prepare 1x20ml of SpC media in a 200ml flask (autoclaved in the morning)
  • Prepare aliquots of SpII media (autoclaved in the morning)
Wednesday
  • Prepare competent cells using the transformation 1 protocol,
  • Electroporate the compete cells from the transformation protocol 2 using a range of concentrations
Thursday
  • Transform the competent cells prepared from transformation 2 protocol
  • Check the transformation cells from protocol 2, if transformed correctly then carry the test for correct integration,
Friday
  • If the transformation has been successful then carry out the integration test.

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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