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Team:HKU HongKong/Data/Protocols.html
From 2012.igem.org
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| + | <h2 style="font-variant: normal; vertical-align: baseline; clear: left; color: #232323; font-family: Gentium Basic; letter-spacing: normal; line-height: 19.5px; orphans: 2; text-align: justify; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; border: 0px none; margin: 0.7em 0px; padding: 0px"> | ||
| + | <span style="font-weight: 400"><font face="Trebuchet MS" size="6"> | ||
| + | Molecular Cloning Protocols</font></span></h2> | ||
| + | <p style="font-variant: normal; vertical-align: baseline; clear: left; color: #232323; font-family: Gentium Basic; letter-spacing: normal; line-height: 19.5px; orphans: 2; text-align: justify; text-indent: 0px; text-transform: none; white-space: normal; widows: 2; word-spacing: 0px; -webkit-text-size-adjust: auto; -webkit-text-stroke-width: 0px; border: 0px none; margin: 0.7em 0px; padding: 0px"> | ||
| + | <br> | ||
| + | <b> | ||
| + | <span style="font-family: Trebuchet MS; font-weight: 400; text-decoration: underline"> | ||
| + | <i><font size="4" color="#232323">Transforming Competent E.coli with the | ||
| + | Plasmid: <br> | ||
| + | <br> | ||
| + | </font></i></span></b><font size="2" face="Tahoma">We chose the biobrick K137076 to | ||
| + | ligate to the pvdQ gene for the following reasons: </font></font></p> | ||
| + | <ul> | ||
| + | <li> | ||
| + | <p style="text-align: justify"> | ||
| + | <font face="Tahoma" size="2" color="#232323">Take out the competent E-coli | ||
| + | cells from the -80 freezer. (Keep all tubes on ice).<br> | ||
| + | </font></li> | ||
| + | <li> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> | ||
| + | <font color="#232323"> | ||
| + | <span style="font-size: 10pt; font-family: Tahoma; font-weight: 400">Add 1uL | ||
| + | of the plasmid DNA in 100uL competent cells (if from Kit). For | ||
| + | transformation of ligated product, add 10uL of the ligated plasmid DNA in | ||
| + | 100uL competent cells. </span></font></p> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> | ||
| + | <font color="#232323"> | ||
| + | <span style="font-size: 10pt; font-family: Tahoma; font-weight: 400"> | ||
| + | Incubate on ice for 10 min. </span></font></li> | ||
| + | </ul> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> </p> | ||
| + | <ul> | ||
| + | <li> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> | ||
| + | <font color="#232323"> | ||
| + | <span style="font-size: 10pt; font-family: Tahoma; font-weight: 400">Place | ||
| + | in water bath at 42°C for 90s.</span></font></li> | ||
| + | </ul> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> </p> | ||
| + | <ul> | ||
| + | <li> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> | ||
| + | <font color="#232323"> | ||
| + | <span style="font-size: 10pt; font-family: Tahoma; font-weight: 400">Place | ||
| + | immediately back on ice for at least 2 min</span></font></li> | ||
| + | </ul> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> </p> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> </p> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> </p> | ||
| + | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> </p> | ||
| + | <p style="text-align: justify"> | ||
</body> | </body> | ||
</html> | </html> | ||
Revision as of 14:07, 23 September 2012
Team:HKU HK
From 2011.igem.org
Molecular Cloning Protocols
Transforming Competent E.coli with the
Plasmid:
We chose the biobrick K137076 to
ligate to the pvdQ gene for the following reasons:
-
Take out the competent E-coli cells from the -80 freezer. (Keep all tubes on ice).
-
Add 1uL of the plasmid DNA in 100uL competent cells (if from Kit). For transformation of ligated product, add 10uL of the ligated plasmid DNA in 100uL competent cells.
Incubate on ice for 10 min.
-
Place in water bath at 42°C for 90s.
-
Place immediately back on ice for at least 2 min
