INSA-Lyon/23 September 2010

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September 23rd

DNA extraction of I0500, 4O, 10L, 8C/14K (2 clones) ; Agarose gel electrophoresis for verification :
There is no more plasmide for 4O, 10L and I0500. We will ask some new.


Transformation of the ligation overnight of yesterday


Measure of fluorescence and DO of the liquid culture of PHL1273 of the 20/09. Start of 3 new cultures in a waterbath at 37°C with a shaking speed of 100 rpm



Competent cells production of PHL818 strain: 500µl overnight culture in 100ml LB-Tet, at 37°C until DO600nm=0,49. Chemiocompetence had been done with protocol described on wiki. 12 competent bacteria vials was stored at -80°C.


4 plasmid DNA extractions (14K (Amp), I0500 (Kan), 1F/24A (Tet) and 18A/24A (Cm)) with NucleoSpin Plasmid kit (Macherey-Nagel) with four different antibiotic resistances. Goal of the experiment: production of plasmid backbones.


Lyophilisated primers SB-prep-3P and 2Ea are diluted in sterile water (at 100µM): 3P with 325µl and 2Ea with 356µl.


5 PCR had been done in order to produce iGEM backbones: vial 1 corresponded to the negative control with 10µl sterile water, vial 2 corresponded to the plasmid Amp, vial 3 corresponded to the plasmid Kan, vial 4 corresponded to the plasmid Tet and vial 5 corresponded to the plasmid Cm.


MIX composition for one PCR:


  1. 5µl 10X buffer with MgCl2
  2. QSP H2O
  3. 5µl dNTP (PCR Dig Labeling Mix, Roche)
  4. 10µl plasmid DNA (diluted 1/5)
  5. 1µl of each primer
  6. 0,75µl Taq Pol (Euromedex)

NB: Hybridation temperature : 58°C (program PSB1)


Gel verification (0,7%) without ladder (no more!!): Negative control OK, Amp: too many strips (no strip at the expected size), Kan: no DNA (I0500 plasmid extraction never gives DNA, it’s confirmed), Tet et Cm: the two biggest strips seem at the good sizes (2,5 and 2kb). PCR products and diluted primers are stored at -20°C.








June
MTWTFSS
  [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/1_June_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/2_June_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/3_June_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/4_June_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/5_June_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/6_June_2010&action=edit 6]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/7_June_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/8_June_2010&action=edit 8] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/9_June_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/10_June_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/11_June_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/12_June_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/13_June_2010&action=edit 13]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/14_June_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/15_June_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/16_June_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/17_June_2010&action=edit 17] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/18_June_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/19_June_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/20_June_2010&action=edit 20]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/21_June_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/22_June_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/23_June_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/24_June_2010&action=edit 24] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/25_June_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/26_June_2010&action=edit 26] [http://2010.igem.org/INSA-Lyon/27_June_2010 27]
[http://2010.igem.org/INSA-Lyon/28_June_2010 28] [http://2010.igem.org/INSA-Lyon/29_June_2010 29] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/30_June_2010&action=edit 30]
July
MTWTFSS
      [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/1_July_2010&action=edit 1] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/2_July_2010&action=edit 2] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/3_July_2010&action=edit 3] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/4_July_2010&action=edit 4]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/5_July_2010&action=edit 5] [http://2010.igem.org/INSA-Lyon/6_July_2010 6] [http://2010.igem.org/INSA-Lyon/7_July_2010 7] [http://2010.igem.org/INSA-Lyon/8_July_2010 8] [http://2010.igem.org/INSA-Lyon/9_July_2010 9] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/10_July_2010&action=edit 10] [http://2010.igem.org/INSA-Lyon/11_July_2010 11]
[http://2010.igem.org/INSA-Lyon/12_July_2010 12] [http://2010.igem.org/INSA-Lyon/13_July_2010 13] [http://2010.igem.org/INSA-Lyon/14_July_2010 14] [http://2010.igem.org/INSA-Lyon/15_July_2010 15] [http://2010.igem.org/INSA-Lyon/16_July_2010 16] [http://2010.igem.org/INSA-Lyon/17_July_2010 17] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/18_July_2010&action=edit 18]
[http://2010.igem.org/INSA-Lyon/19_July_2010 19] [http://2010.igem.org/INSA-Lyon/20_July_2010 20] [http://2010.igem.org/INSA-Lyon/21_July_2010 21] [http://2010.igem.org/INSA-Lyon/22_July_2010 22] [http://2010.igem.org/INSA-Lyon/23_July_2010 23] [http://2010.igem.org/INSA-Lyon/24_July_2010 24] [http://2010.igem.org/INSA-Lyon/25_July_2010 25]
[http://2010.igem.org/INSA-Lyon/26_July_2010 26] [http://2010.igem.org/INSA-Lyon/27_July_2010 27] [http://2010.igem.org/INSA-Lyon/28_July_2010 28] [http://2010.igem.org/INSA-Lyon/29_July_2010 29] [http://2010.igem.org/INSA-Lyon/30_July_2010 30] [http://2010.igem.org/INSA-Lyon/31_July_2010 31]
August
MTWTFSS
            [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/1_August_2010&action=edit 1]
[http://2010.igem.org/INSA-Lyon/2_August_2010 2] [http://2010.igem.org/INSA-Lyon/3_August_2010 3] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/4_August_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/5_August_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/6_August_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/7_August_2010&action=edit 7] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/8_August_2010&action=edit 8]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/9_August_2010&action=edit 9] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/10_August_2010&action=edit 10] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/11_August_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/12_August_2010&action=edit 12] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/13_August_2010&action=edit 13] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/14_August_2010&action=edit 14] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/15_August_2010&action=edit 15]
[http://2010.igem.org/INSA-Lyon/16_August_2010 16] [http://2010.igem.org/INSA-Lyon/17_August_2010 17] [http://2010.igem.org/INSA-Lyon/18_August_2010 18] [http://2010.igem.org/INSA-Lyon/19_August_2010 19] [http://2010.igem.org/INSA-Lyon/20_August_2010 20] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/21_August_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/22_August_2010&action=edit 22]
[http://2010.igem.org/INSA-Lyon/23_August_2010 23] [http://2010.igem.org/INSA-Lyon/24_August_2010 24] [http://2010.igem.org/INSA-Lyon/25_August_2010 25] [http://2010.igem.org/INSA-Lyon/26_August_2010 26] [http://2010.igem.org/INSA-Lyon/27_August_2010 27] [http://2010.igem.org/INSA-Lyon/28_August_2010 28] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/29_August_2010&action=edit 29]
[http://2010.igem.org/INSA-Lyon/30_August_2010 30] [http://2010.igem.org/INSA-Lyon/31_August_2010 31]
September
MTWTFSS
    [http://2010.igem.org/INSA-Lyon/1_September_2010 1] [http://2010.igem.org/INSA-Lyon/2_September_2010 2] [http://2010.igem.org/INSA-Lyon/3_September_2010 3] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/4_September_2010&action=edit 4] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/5_September_2010&action=edit 5]
[http://2010.igem.org/INSA-Lyon/6_September_2010 6] [http://2010.igem.org/INSA-Lyon/7_September_2010 7] [http://2010.igem.org/INSA-Lyon/8_September_2010 8] [http://2010.igem.org/INSA-Lyon/9_September_2010 9] [http://2010.igem.org/INSA-Lyon/10_September_2010 10] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/11_September_2010&action=edit 11] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/12_September_2010&action=edit 12]
[http://2010.igem.org/INSA-Lyon/13_September_2010 13] [http://2010.igem.org/INSA-Lyon/14_September_2010 14] [http://2010.igem.org/INSA-Lyon/15_September_2010 15] [http://2010.igem.org/INSA-Lyon/16_September_2010 16] [http://2010.igem.org/INSA-Lyon/17_September_2010 17] [http://2010.igem.org/INSA-Lyon/18_September_2010 18] [http://2010.igem.org/INSA-Lyon/19_September_2010 19]
[http://2010.igem.org/INSA-Lyon/20_September_2010 20] [http://2010.igem.org/INSA-Lyon/21_September_2010 21] [http://2010.igem.org/INSA-Lyon/22_September_2010 22] [http://2010.igem.org/INSA-Lyon/23_September_2010 23] [http://2010.igem.org/INSA-Lyon/24_September_2010 24] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/25_September_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/26_September_2010&action=edit 26]
[http://2010.igem.org/INSA-Lyon/27_September_2010 27] [http://2010.igem.org/INSA-Lyon/28_September_2010 28] [http://2010.igem.org/INSA-Lyon/29_September_2010 29] [http://2010.igem.org/INSA-Lyon/30_September_2010 30]
October
MTWTFSS
        [http://2010.igem.org/INSA-Lyon/1_October_2010 1] [http://2010.igem.org/INSA-Lyon/2_October_2010 2] [http://2010.igem.org/INSA-Lyon/3_October_2010 3]
[http://2010.igem.org/INSA-Lyon/4_October_2010 4] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/5_October_2010&action=edit 5] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/6_October_2010&action=edit 6] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/7_October_2010&action=edit 7] [http://2010.igem.org/INSA-Lyon/8_October_2010 8] [http://2010.igem.org/INSA-Lyon/9_October_2010 9] [http://2010.igem.org/INSA-Lyon/10_October_2010 10]
[http://2010.igem.org/INSA-Lyon/11_October_2010 11] [http://2010.igem.org/INSA-Lyon/12_October_2010 12] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/13_October_2010&action=edit 13] [http://2010.igem.org/INSA-Lyon/14_October_2010 14] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/15_October_2010&action=edit 15] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/16_October_2010&action=edit 16] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/17_October_2010&action=edit 17]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/18_October_2010&action=edit 18] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/19_October_2010&action=edit 19] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/20_October_2010&action=edit 20] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/21_October_2010&action=edit 21] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/22_October_2010&action=edit 22] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/23_October_2010&action=edit 23] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/24_October_2010&action=edit 24]
[http://2010.igem.org/wiki/index.php?title=INSA-Lyon/25_October_2010&action=edit 25] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/26_October_2010&action=edit 26] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/27_October_2010&action=edit 27] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/28_October_2010&action=edit 28] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/29_October_2010&action=edit 29] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/30_October_2010&action=edit 30] [http://2010.igem.org/wiki/index.php?title=INSA-Lyon/31_October_2010&action=edit 31]

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