Difference between revisions of "Team:Goettingen/Experiments"
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− | #menu1, #menu2, #menu3, #menu4, #menu5, #menu6, #menu7, #menu8, #menu9, #menu10, #menu11, #menu12, #menu13, #menu14, #menu15, #menu16, #menu17, #menu18{ | + | #menu1, #menu2, #menu3, #menu4, #menu5, #menu6, #menu7, #menu8, #menu9, #menu10, #menu11, #menu12, #menu13, #menu14, #menu15, #menu16, #menu17, #menu18, #menu19, #menu20{ |
display:none; | display:none; | ||
width:90%; | width:90%; | ||
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</ul> | </ul> | ||
</div> | </div> | ||
+ | |||
+ | <a href="" onClick=" $('#menu19').slideToggle(300, function callback() { }); return false;"><h1> Restriction control using fast and slow digest enzymes</h1></a> | ||
+ | <div id="menu19"> | ||
+ | <p align="center"><strong>Restriction control using fast digest enzymes</strong></p> | ||
+ | <p><strong> </strong></p> | ||
+ | <p><strong>Sac I and HindIII - thermo fisher scientific - restriction control protocol</strong> </p> | ||
+ | <p> </p> | ||
+ | <ul> | ||
+ | <li>Add the following components </li> | ||
+ | </ul> | ||
+ | <table style="width: 223px;" border="1" cellspacing="0" cellpadding="0" align="left"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top" width="127"> | ||
+ | <p>Purified Plasmid </p> | ||
+ | </td> | ||
+ | <td valign="top" width="96"> | ||
+ | <p>500-1000 ng</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="127"> | ||
+ | <p>Sac I </p> | ||
+ | </td> | ||
+ | <td valign="top" width="96"> | ||
+ | <p>1 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="127"> | ||
+ | <p>Hind III </p> | ||
+ | </td> | ||
+ | <td valign="top" width="96"> | ||
+ | <p>1 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="127"> | ||
+ | <p>Nuclease free H<sub>2</sub>O </p> | ||
+ | </td> | ||
+ | <td valign="top" width="96"> | ||
+ | <p>Add to 10 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <ul> | ||
+ | <li>Incubate at 37 <sup>o</sup>C for 30 min.</li> | ||
+ | <li>For inactivation incubation at 65°C for 5 min. </li> | ||
+ | </ul> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p><strong>PST I - thermo fisher scientific - restriction control protocol</strong> </p> | ||
+ | <p> </p> | ||
+ | <ul> | ||
+ | <li>Add the following components </li> | ||
+ | </ul> | ||
+ | <table style="width: 254px;" border="1" cellspacing="0" cellpadding="0" align="left"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>Purified Plasmid </p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>500-1000 ng</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>FD buffer</p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>1 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>PST I </p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>1 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>Nuclease free H<sub>2</sub>O </p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>Add to 10 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <ul> | ||
+ | <li>Incubate at 37 <sup>o</sup>C for 30 min.</li> | ||
+ | <li>Products were checked on 0.8% agarose gel.</li> | ||
+ | </ul> | ||
+ | <p> </p> | ||
+ | <p align="center"><strong>Restriction control using slow digest enzymes</strong></p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p><strong>Bsm I - fermentas - restriction control protocol</strong> </p> | ||
+ | <p> </p> | ||
+ | <ul> | ||
+ | <li>Add the following components </li> | ||
+ | </ul> | ||
+ | <table style="width: 254px;" border="1" cellspacing="0" cellpadding="0" align="left"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>Purified Plasmid </p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>500 ng </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>Red buffer</p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>1 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>Bsm I </p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>1 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top"> | ||
+ | <p>Nuclease free H<sub>2</sub>O </p> | ||
+ | </td> | ||
+ | <td valign="top"> | ||
+ | <p>Add to 8 µl </p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <p> </p> | ||
+ | <ul> | ||
+ | <li>Incubate at 37 <sup>o</sup>C for 3 hours.</li> | ||
+ | <li>Products were checked on 0.8% agarose gel.</li> | ||
+ | </ul> | ||
+ | <p> </p> | ||
+ | <p></p> | ||
+ | <p> </p> | ||
+ | <p></p> | ||
+ | <p> </p> | ||
+ | </div> | ||
+ | |||
+ | |||
+ | <a href="" onClick=" $('#menu20').slideToggle(300, function callback() { }); return false;"> | ||
+ | <h1> Scafoldin Restriction control </h1></a> | ||
+ | <div id="menu20"> | ||
+ | |||
+ | <p align="center"><span style="font-size: x-large; color: #888888;"><strong></strong></span></p> | ||
+ | <p align="center"> </p> | ||
+ | <ul> | ||
+ | <p><strong>Scaffoldin Restriction Control using the restriction enzyme BsmI</strong></p> | ||
+ | <p>Set up the reaction as follows,</p> | ||
+ | <table border="1" cellspacing="0" cellpadding="0"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>Components</p> | ||
+ | </td> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>Quantity</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>Nuclease free Water</p> | ||
+ | </td> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>15 µl</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>Red Buffer 10X</p> | ||
+ | </td> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>2 µl</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>DNA</p> | ||
+ | </td> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>1000 ng (2 µl)</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>BsmI</p> | ||
+ | </td> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>1µl</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" width="301"> | ||
+ | <p> </p> | ||
+ | </td> | ||
+ | <td valign="top" width="301"> | ||
+ | <p>20 µl</p> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <p> </p> | ||
+ | <p>Incubate the mixture for 2 hours at 37ᵒC.</p> | ||
+ | <p>Perform the heat inactivation step for the reaction mixture for 10 min at 65ᵒC.</p> | ||
+ | </div> | ||
+ | |||
</html> | </html> |
Revision as of 14:46, 15 September 2015