Team:UNIPV-Pavia/Calendar/July/settimana5

From 2011.igem.org

(Difference between revisions)
Line 87: Line 87:
<p>
<p>
-
Digestions of previously purified plasmids (carrying either E17, or E18, or E19 or E20 part) were performed for ligations:
+
Reactions were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols.
</p>
</p>
 +
 +
<p>
 +
In the afternoon gel electrophoresis was performed.
 +
</p>
 +
 +
<p>
 +
Immagine della corsa su gel
 +
</p>
 +
 +
<p>
 +
After gel extraction, digested DNA was quantified:
 +
</p>
 +
 +
<center>
<center>
<table border="1">
<table border="1">
     <tr>
     <tr>
-
       <td><b>Plasmid</b></td>
+
       <td><b>Part</b></td>
-
      <td><b>Kind</b></td>
+
       <td><b>DNA (ng/&mu;l)</b></td>
-
       <td><b>DNA (&mu;l)</b></td>
+
-
      <td><b>H<small><sub>2</sub></small>O (&mu;l)</b></td>
+
-
      <td><b>Enzyme 1 (&mu;l)</b></td>
+
-
      <td><b>Enzyme 2 (&mu;l)</b></td>
+
-
      <td><b>Buffer H (&mu;l)</b></td>
+
-
      <td><b>Final Volume (&mu;l)</b></td>
+
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E17</td>
+
       <td>E24 (E-P)</td>
-
      <td>Insert</td>
+
       <td>-</td>
       <td>-</td>
-
      <td>-</td>
 
-
      <td>1 Xbal</td>
 
-
      <td>1 Pstl</td>
 
-
      <td>2.5</td>
 
-
      <td>25</td>
 
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E18</td>
+
       <td>E25 (E-P)</td>
-
      <td>Insert</td>
+
       <td>-</td>
       <td>-</td>
-
      <td>-</td>
 
-
      <td>1 Xbal</td>
 
-
      <td>1 Pstl</td>
 
-
      <td>2.5</td>
 
-
      <td>25</td>
 
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E19</td>
+
       <td>E26 (E-P)</td>
-
      <td>Insert</td>
+
-
      <td>-</td>
+
       <td>-</td>
       <td>-</td>
-
      <td>1 Xbal</td>
 
-
      <td>1 Pstl</td>
 
-
      <td>2.5</td>
 
-
      <td>25</td>
 
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E20</td>
+
       <td>E27 (E-P)</td>
-
      <td>Insert</td>
+
       <td>-</td>
       <td>-</td>
-
      <td>-</td>
 
-
      <td>1 Xbal</td>
 
-
      <td>1 PstI</td>
 
-
      <td>2.5</td>
 
-
      <td>25</td>
 
   </tr>
   </tr>
Line 154: Line 138:
<p>
<p>
-
Reactions were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols.
+
Previously purified plasmids carrying either E17, or E18, or E19 or E20 part were inoculated in order to amplify them for the subsequent transformation in MGZ1 cells (E' GIUSTO?):
-
</p>
+
-
 
+
-
<p>
+
-
In the afternoon gel electrophoresis was performed.
+
</p>
</p>
 +
<p><a name="indice"/>
 +
</p>
 +
<a name="July.2C_26th"></a><h2> <span class="mw-headline">July, 26th</span></h2>
<p>
<p>
-
Immagine della corsa su gel
 
-
</p>
 
<p>
<p>
-
After gel extraction, digested DNA was quantified:
+
Plasmids containing either E17, or E18, or E19, or E20 or E36 part were extracted with MiniPrep kit from ON saturated cultures; purified DNA was quantified with NanoDrop Spectrophotometer:
</p>
</p>
-
 
-
 
<center>
<center>
<table border="1">
<table border="1">
     <tr>
     <tr>
-
       <td><b>Part</b></td>
+
       <td><b>Plasmid</b></td>
       <td><b>DNA (ng/&mu;l)</b></td>
       <td><b>DNA (ng/&mu;l)</b></td>
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E24 (E-P)</td>
+
       <td>E17</td>
-
       <td>-</td>
+
       <td>20.3</td>
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E25 (E-P)</td>
+
       <td>E18</td>
-
       <td>-</td>
+
       <td>18.6</td>
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E26 (E-P)</td>
+
       <td>E19</td>
-
       <td>-</td>
+
       <td>17.3</td>
   </tr>
   </tr>
   <tr>
   <tr>
-
       <td>E27 (E-P)</td>
+
       <td>E20</td>
-
       <td>-</td>
+
       <td>13.2</td>
 +
  </tr>
 +
 
 +
  <tr>
 +
      <td>E36</td>
 +
      <td>17.8</td>
   </tr>
   </tr>
</table>
</table>
</center>
</center>
 +
<p>
 +
Digestion of E36 part carrying plasmid was performed for ligation:
 +
</p>
 +
 +
<center>
 +
<table border="1">
 +
    <tr>
 +
      <td><b>Plasmid</b></td>
 +
      <td><b>Kind</b></td>
 +
      <td><b>DNA (&mu;l)</b></td>
 +
      <td><b>H<small><sub>2</sub></small>O (&mu;l)</b></td>
 +
      <td><b>Enzyme 1 (&mu;l)</b></td>
 +
      <td><b>Enzyme 2 (&mu;l)</b></td>
 +
      <td><b>Buffer H (&mu;l)</b></td>
 +
      <td><b>Final Volume (&mu;l)</b></td>
 +
  </tr>
 +
 +
  <tr>
 +
      <td>E36</td>
 +
      <td>Insert</td>
 +
      <td>20.5</td>
 +
      <td>0</td>
 +
      <td>1 Xbal</td>
 +
      <td>1 Pstl</td>
 +
      <td>2.5</td>
 +
      <td>25</td>
 +
  </tr>
 +
 +
</table>
 +
</center>
 +
 +
<table border="0" width="100%" class="menu">
<table border="0" width="100%" class="menu">

Revision as of 20:26, 28 July 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

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

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) -

Previously purified plasmids carrying either E17, or E18, or E19 or E20 part were inoculated in order to amplify them for the subsequent transformation in MGZ1 cells (E' GIUSTO?):

July, 26th

Plasmids containing either E17, or E18, or E19, or E20 or E36 part were extracted with MiniPrep kit from ON saturated cultures; purified DNA was quantified with NanoDrop Spectrophotometer:

Plasmid DNA (ng/μl)
E17 20.3
E18 18.6
E19 17.3
E20 13.2
E36 17.8

Digestion of E36 part carrying plasmid was performed for ligation:

Plasmid Kind DNA (μl) H2O (μl) Enzyme 1 (μl) Enzyme 2 (μl) Buffer H (μl) Final Volume (μl)
E36 Insert 20.5 0 1 Xbal 1 Pstl 2.5 25
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