Team:SDU-Denmark/labnotes9

From 2010.igem.org

(Difference between revisions)
(Photosensor group)
(Insertion of PS in pSB1C3 and pSB1AK3)
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'''Date:''' 9/9 2010<Br>
'''Date:''' 9/9 2010<Br>
'''Done By:''' Maria and Lc<Br>
'''Done By:''' Maria and Lc<Br>
-
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1 CP1.1]<br>
+
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#DE1.3 DE1.3]<br>
'''Notes:'''<br>
'''Notes:'''<br>
 +
DNA was extracted from gel according to protocol and each sample was diluted in 20uL.<br><br>
 +
'''Analysis:'''<br>
 +
each sample had a concentration of app. 7ng/uL. All 4 samples were pooled and used for restriction digest.<br><br>
 +
 +
==== Restriction digest of PS, pSB1C3 and pSB1AK3 ====
 +
'''Date:''' 9/9 2010<Br>
 +
'''Done By:''' Maria and Lc<Br>
 +
'''Protocol:'''[https://2010.igem.org/Team:SDU-Denmark/protocols#RD1.1 RD1.1][https://2010.igem.org/Team:SDU-Denmark/protocols#DE1.3 DE1.3]<br>
 +
'''Notes:'''<br>
 +
Restriction mixture pSB1C3:<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>H<small>2</small>O</td>
 +
      <td>24uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>FD green buffer</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>EcoRI</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PstI</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB1C3</td>
 +
      <td>10uL</td>
 +
    </tr>
 +
</table><br><br>
 +
 +
Restriction mixture pSB1AK3:<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>H<small>2</small>O</td>
 +
      <td>24uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>FD green buffer</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>EcoRI</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>XbaI</td>
 +
      <td>2uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB1AK3</td>
 +
      <td>10uL</td>
 +
    </tr>
 +
</table><br><br>
 +
 +
Restriction mixture PS (used in pSB1C3):<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>H<small>2</small>O</td>
 +
      <td>38uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>FD green buffer</td>
 +
      <td>8uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>EcoRI</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PstI</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PS</td>
 +
      <td>30uL</td>
 +
    </tr>
 +
</table><br><br>
 +
 +
Restriction mixture PS (used in pSB1AK3):<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>H<small>2</small>O</td>
 +
      <td>38uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>FD green buffer</td>
 +
      <td>8uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>EcoRI</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>SpeI</td>
 +
      <td>4uL</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PS</td>
 +
      <td>30uL</td>
 +
    </tr>
 +
</table><br><br>
 +
 +
Digested samples were loaded onto a 1.5% agarose extraction gel. Uncut PS, pSB1C3 and pSB1AK3 were used as controles. Gene ruler DNA ladder mix was used as marker.<br>
 +
Loading scheme:<br>
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>Lane</td>
 +
      <td>sample</td>
 +
    </tr>
 +
    <tr>
 +
      <td>1</td>
 +
      <td>marker</td>
 +
    </tr>
 +
    <tr>
 +
      <td>2</td>
 +
      <td>PS (used in pSB1C3)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>3</td>
 +
      <td>uncut PS</td>
 +
    </tr>
 +
    <tr>
 +
      <td>4</td>
 +
      <td>PS (used in pSB1AK3)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>5</td>
 +
      <td>uncut pSB1C3</td>
 +
    </tr>
 +
    <tr>
 +
      <td>6</td>
 +
      <td>pSB1C3</td>
 +
    </tr>
 +
    <tr>
 +
      <td>7</td>
 +
      <td>uncut pSB1AK3</td>
 +
    </tr>
 +
    <tr>
 +
      <td>8</td>
 +
      <td>pSB1AK3</td>
 +
    </tr>
 +
</table><br>
 +
DNA was extracted from gel according to protocol.<br><br>
 +
'''Results:'''<br>
 +
DNA conc:
 +
<table style="text-align: left;" border="1"
 +
cellpadding="2" cellspacing="2">
 +
    <tr>
 +
      <td>sample</td>
 +
      <td>conc. (ng/uL)</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PS (used in pSB1C3)</td>
 +
      <td>5.5</td>
 +
    </tr>
 +
    <tr>
 +
      <td>PS (used in pSB1AK3)</td>
 +
      <td>4.2</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB1C3</td>
 +
      <td>6.3</td>
 +
    </tr>
 +
    <tr>
 +
      <td>pSB1AK3</td>
 +
      <td>14.3</td>
 +
    </tr>
 +
</table><br><br>
 +
'''Analysis:'''
 +
the purified DNA was used for ligation.<br><br>

Revision as of 13:08, 21 September 2010