Team:UNIPV-Pavia/Calendar/July/settimana5

From 2011.igem.org

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Digestions of previously purified plasmids (carrying either E17, or E18, or E19 or E20 part) were performed for ligations:
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<table border="1">
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    <tr>
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      <td><b>Plasmid</b></td>
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      <td><b>Kind</b></td>
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      <td><b>DNA (&mu;l)</b></td>
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      <td><b>H<small><sub>2</sub></small>O (&mu;l)</b></td>
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      <td><b>Enzyme 1 (&mu;l)</b></td>
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      <td><b>Enzyme 2 (&mu;l)</b></td>
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      <td><b>Buffer H (&mu;l)</b></td>
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      <td><b>Final Volume (&mu;l)</b></td>
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  </tr>
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  <tr>
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      <td>E17</td>
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      <td>Insert</td>
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      <td>-</td>
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      <td>-</td>
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      <td>1 Xbal</td>
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      <td>1 Pstl</td>
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      <td>2.5</td>
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      <td>25</td>
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  </tr>
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  <tr>
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      <td>E18</td>
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      <td>Insert</td>
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      <td>-</td>
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      <td>-</td>
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      <td>1 Xbal</td>
 +
      <td>1 Pstl</td>
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      <td>2.5</td>
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      <td>25</td>
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  </tr>
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  <tr>
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      <td>E19</td>
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      <td>Insert</td>
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      <td>-</td>
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      <td>-</td>
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      <td>1 Xbal</td>
 +
      <td>1 Pstl</td>
 +
      <td>2.5</td>
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      <td>25</td>
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  </tr>
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 +
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  <tr>
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      <td>E20</td>
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      <td>Insert</td>
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      <td>-</td>
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      <td>-</td>
 +
      <td>1 Xbal</td>
 +
      <td>1 PstI</td>
 +
      <td>2.5</td>
 +
      <td>25</td>
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  </tr>
</table>
</table>

Revision as of 19:55, 28 July 2011

UNIPV TEAM 2011

March
M T W T F S S
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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
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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

JULY: WEEK 5

July, 25th

Red colonies were observed from E36 plates, suggesting that the transformation was successful. A colony was picked form the plate.

Digestions of previously purified plasmids were performed for ligations:

Plasmid Kind DNA (μl) H2O (μl) Enzyme 1 (μl) Enzyme 2 (μl) Buffer H (μl) Final Volume (μl)
E24-2 Insert 13 7.5 1 Xbal 1 Pstl 2.5 25
E25-1 Insert 13.5 7 1 Xbal 1 Pstl 2.5 25
E26-2 Insert 14.5 6 1 Xbal 1 Pstl 2.5 25
E27-1 Insert 12.5 8 1 Xbal 1 PstI 2.5 25

Digestions of previously purified plasmids (carrying either E17, or E18, or E19 or E20 part) were performed for ligations:

Plasmid Kind DNA (μl) H2O (μl) Enzyme 1 (μl) Enzyme 2 (μl) Buffer H (μl) Final Volume (μl)
E17 Insert - - 1 Xbal 1 Pstl 2.5 25
E18 Insert - - 1 Xbal 1 Pstl 2.5 25
E19 Insert - - 1 Xbal 1 Pstl 2.5 25
E20 Insert - - 1 Xbal 1 PstI 2.5 25

Reactions were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols.

In the afternoon gel electrophoresis was performed.

Immagine della corsa su gel

After gel extraction, digested DNA was quantified:

Part DNA (ng/μl)
E24 (E-P) -
E25 (E-P) -
E26 (E-P) -
E27 (E-P) -