INSA-Lyon/20 September 2010

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<li>waterbath at 31°C, shaking speed 100 rpm</li>
<li>waterbath at 31°C, shaking speed 100 rpm</li>
<li>waterbath at 32°C, shaking speed 100 rpm </li>
<li>waterbath at 32°C, shaking speed 100 rpm </li>
 +
</ol><br></p><br>
 +
<p>Lyophilisated primers ompR234 Forward and Reverse are diluted in sterile water (at 100µM): Forward primer: 450µl added, and Reverse primer: 560µl added.</p><br>
 +
<p>3 PCR: vial 1 corresponded to negative control with 10µl sterile water, vial 2 corresponded to genomic DNA 1/5 diluted and vial 3 corresponded to genomic DNA 1/10 diluted.</p><br>
 +
<p>MIX composition for one PCR:<br>
 +
<ol id=”list_extra”>
 +
<li>5µl 10X buffer with MgCl2</li>
 +
<li>QSP H2O</li>
 +
<li>5µl dNTP (PCR Dig Labeling Mix, Roche) </li>
 +
<li>10 or 5 µl DNA</li>
 +
<li>1µl of each primer</li>
 +
<li>0,75µl Taq Pol (Euromedex)</li>
</ol><br></p>
</ol><br></p>
 +
<p>NB: Hybridation temperature is 56°C (program OMPR). PCR products and diluted primers are always stored at -20°C.</p>
<br><br><br><br><br><br><br>
<br><br><br><br><br><br><br>

Revision as of 07:20, 6 October 2010






September 20th


Digestion with E+P of :

  1. 1F/24A (3 clones)
  2. 4O/10L
  3. Curli/22B (2 clones)
  4. I0500 just with E (clone from the 13/08 and the 17/08


Verification of the weight of the part and the ligation on an agarose gel electrophoris :

  1. I0500, 4O/10L : the part are not correct, we will ask for I0500, 4O and 10L again
  2. One clone of Curli/22B and all the 1F/24A seams OK

Start of 6 liquid cultur of PHL1273 with 2 condition of cultur :

  1. waterbath at 31°C, shaking speed 100 rpm
  2. waterbath at 32°C, shaking speed 100 rpm


Lyophilisated primers ompR234 Forward and Reverse are diluted in sterile water (at 100µM): Forward primer: 450µl added, and Reverse primer: 560µl added.


3 PCR: vial 1 corresponded to negative control with 10µl sterile water, vial 2 corresponded to genomic DNA 1/5 diluted and vial 3 corresponded to genomic DNA 1/10 diluted.


MIX composition for one PCR:

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

NB: Hybridation temperature is 56°C (program OMPR). PCR products and diluted primers are always 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]