Team:SDU-Denmark/labnotes9
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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# | + | '''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