Difference between revisions of "Team:UrbanTundra Edmonton/Notebook"
m |
m |
||
| Line 3: | Line 3: | ||
<div class="column full_size"> | <div class="column full_size"> | ||
| − | + | <b>July 20th, 2016</b> | |
<ol>4 microlitres of milliq water with 1 microlitre of plasmid</ol> | <ol>4 microlitres of milliq water with 1 microlitre of plasmid</ol> | ||
<ol>5 microlitres of no chlorite dismutase</ol> | <ol>5 microlitres of no chlorite dismutase</ol> | ||
| Line 14: | Line 14: | ||
<ol>Add 2 microlitres of T4DNA ligase buffer. </ol> | <ol>Add 2 microlitres of T4DNA ligase buffer. </ol> | ||
<ol>Add 0.5 microlitre of ligase then mix gently and incubate at room temperature for 30 minutes or longer. </ol> | <ol>Add 0.5 microlitre of ligase then mix gently and incubate at room temperature for 30 minutes or longer. </ol> | ||
| − | + | ||
</div> | </div> | ||
</html> | </html> | ||
Revision as of 06:24, 16 October 2016
July 20th, 2016
- 4 microlitres of milliq water with 1 microlitre of plasmid
- 5 microlitres of no chlorite dismutase
- 1 microlitre of cutsmart buffer (specifically 1.1111 microlitre)
- 10 x cutsmart buffer into tubes makes 1x [ ]
- When adding BsaI, only touch the surface of the BsaI to the pipette tip. The reason: has glycerol which is sticky. Take 0.5 microlitre of BsaI.
- Mix sample very gently then put into a 37 degrees celsius incubator for 1 hour.
- Heat kill at 65 degrees celsius for 20 minutes. This kills the BsaI to prevent star activity.
- Pulse down tubes then add 8 microlitre of milliq water.
- Add 2 microlitres of T4DNA ligase buffer.
- Add 0.5 microlitre of ligase then mix gently and incubate at room temperature for 30 minutes or longer.
