Team:UNIPV-Pavia/Calendar/August/week3

From 2011.igem.org

(Difference between revisions)
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Inoculum of E41N-1 and J101-4C5 for plasmid purification 6 ml of LB + Cm12.5.
Inoculum of E41N-1 and J101-4C5 for plasmid purification 6 ml of LB + Cm12.5.
<br>
<br>
-
Streak of E32, E33, J101-31, J101-E5 and ENTERO4C5 to characterize PtetR promoter with different Ribosome Binding Sites (<a href="http://partsregistry.org/wiki/index.php/Part:BBa_B0030">BBa_B0030</a>, <a href="http://partsregistry.org/wiki/index.php/Part:BBa_B0031">BBa_B0031</a>).
+
Streak of E32, E33, J101-31, J101-E5 and ENTERO4C5 on an LB-agar plate + Cm12.5 to test and characterize PtetR promoter with different Ribosome Binding Sites (<a href="http://partsregistry.org/wiki/index.php/Part:BBa_B0030">BBa_B0030</a>, <a href="http://partsregistry.org/wiki/index.php/Part:BBa_B0031">BBa_B0031</a>).
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</p>
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<p>
<p>
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E41N-1 and J101-4C5 plasmids were ourified with mini Prep kit:
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E41N-1 and J101-4C5 plasmids were purified with mini Prep kit:
</p>
</p>
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<p>
<p>
T9002-ENTERO4C5 and ENTERO-RBS were diluted 1:500 in a final volume and 1 ml of 12 ml M9 + Cm12.5 respectively and grown for 6 hours.
T9002-ENTERO4C5 and ENTERO-RBS were diluted 1:500 in a final volume and 1 ml of 12 ml M9 + Cm12.5 respectively and grown for 6 hours.
 +
<br>
 +
3OC6-HSL was diluted to known concentrations in order to induce T9002-ENTERO4C5 biosensor and obtain a reference curve for the measurements on E37, E38, E39, E40 and ENTERO4C5 supernatants.
 +
Supernatants were diluted in M9 + Amp + Cm12.5 in order to obtain 1:500 and 1:200 final dilutions in the 200 &mu;l-well of TECAN microplate.
 +
<br>
 +
In the afternoon supernatants were tested; this time results were more precise, even if negative control seemed to inactivate 3OC<small><sub>6</sub></small>-HSL as fast as other cultures.
 +
<br>
 +
1.5 &mu;l of J101-4C5 purified DNA was transformed in 100 &mu;l of MGZ1 competent cells.
 +
<br>
 +
E32, E33, J101-31, J101-E5 and ENTERO4C5 plate was grown so two colonies for each strain were picked and inoculated in 1 ml of M9 + Cm12.5 for PtetR characterization.
 +
<br>
 +
<a href="http://partsregistry.org/wiki/index.php/Part:BBa_K300005">BBa_K300005</a> was inoculated in 6 ml of LB + Cm34 in order to extract <a href=http://partsregistry.org/Part:pSB1C3>pSB1C3</a> standard shipping vector.
</p>
</p>
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<div align="right"><small><a href="#indice" title="">^top</a></small></div>
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<a name="August.2C_18th"></a><h2> <span class="mw-headline">August, 18th</span></h2>
<div align="right"><small><a href="#indice" title="">^top</a></small></div>
<div align="right"><small><a href="#indice" title="">^top</a></small></div>

Revision as of 20:12, 21 August 2011

UNIPV TEAM 2011

March
M T W T F S S
  1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30 31    

April
M T W T F S S
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30

May
M T W T F S S
            1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31

June
M T W T F S S
    1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30

July
M T W T F S S
        1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31

August
M T W T F S S
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30 31

September
M T W T F S S
      1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30

October
M T W T F S S
          1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31

AUGUST: WEEK 3

August, 16th

Inoculum of T9002-ENTERO4C5 and ENTERO-RBS (1 ml of M9 + Cm12.5) to test again diluted supernatants of E37, E38, E39, E40 and ENTERO4C5.
Inoculum of E41N-1 and J101-4C5 for plasmid purification 6 ml of LB + Cm12.5.
Streak of E32, E33, J101-31, J101-E5 and ENTERO4C5 on an LB-agar plate + Cm12.5 to test and characterize PtetR promoter with different Ribosome Binding Sites (BBa_B0030, BBa_B0031).

August, 17th

E41N-1 and J101-4C5 plasmids were purified with mini Prep kit:

Plasmid DNA (ng/μl)
E41N-1 33.7
J101-4C5 25.4

T9002-ENTERO4C5 and ENTERO-RBS were diluted 1:500 in a final volume and 1 ml of 12 ml M9 + Cm12.5 respectively and grown for 6 hours.
3OC6-HSL was diluted to known concentrations in order to induce T9002-ENTERO4C5 biosensor and obtain a reference curve for the measurements on E37, E38, E39, E40 and ENTERO4C5 supernatants. Supernatants were diluted in M9 + Amp + Cm12.5 in order to obtain 1:500 and 1:200 final dilutions in the 200 μl-well of TECAN microplate.
In the afternoon supernatants were tested; this time results were more precise, even if negative control seemed to inactivate 3OC6-HSL as fast as other cultures.
1.5 μl of J101-4C5 purified DNA was transformed in 100 μl of MGZ1 competent cells.
E32, E33, J101-31, J101-E5 and ENTERO4C5 plate was grown so two colonies for each strain were picked and inoculated in 1 ml of M9 + Cm12.5 for PtetR characterization.
BBa_K300005 was inoculated in 6 ml of LB + Cm34 in order to extract pSB1C3 standard shipping vector.

August, 18th