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Team:HKU HongKong/Data/Protocols.html
From 2012.igem.org
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of the plasmid DNA in 100uL competent cells (if from Kit). For | of the plasmid DNA in 100uL competent cells (if from Kit). For | ||
transformation of ligated product, add 10uL of the ligated plasmid DNA in | transformation of ligated product, add 10uL of the ligated plasmid DNA in | ||
| - | 100uL competent cells. </span></font></p> | + | 100uL competent </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">cells. </span></font></p> | ||
<p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> | <p class="MsoListParagraph" style="text-indent: -18.0pt; text-autospace: none; text-align: left; margin-left: 18.0pt"> | ||
<li> | <li> | ||
Revision as of 14:25, 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.
-
Add 800uL LB broth. Incubate for 1hr at 37°C shaker.
-
Centrifuge at 130rpm for 5min to remove supernatant. Re-suspend the pellet in about 10uL of supernatant.
-
Spread the entire re-suspended pellet on ampicillin agar dishes.
-
Incubate for 12-16 hours at 37 °C.
