Difference between revisions of "Team:Dundee/labjournal manu"
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| − | <p><b>Aim of experiment:</b> | + | |
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| + | <p><b>Aim of experiment:</b>To prepare <a href="http://parts.igem.org/Part:BBa_K1058008">BBa_K1058008</a> for sequence confirmation, storage, and further processing.</p> | ||
| + | |||
<p><b>Protocols Used:</b> | <p><b>Protocols Used:</b> | ||
<p><a data-toggle="modal" data-target="#plating-modal" class="journal-protocol"> Click here to see our plating protocol.</a></p> | <p><a data-toggle="modal" data-target="#plating-modal" class="journal-protocol"> Click here to see our plating protocol.</a></p> | ||
<p><a data-toggle="modal" data-target="#overnightculture-modal" class="journal-protocol"> Click here to see our overnight culture protocol.</a></p></p> | <p><a data-toggle="modal" data-target="#overnightculture-modal" class="journal-protocol"> Click here to see our overnight culture protocol.</a></p></p> | ||
| + | |||
<p><b>Results:</b><a data-toggle="modal" data-target="#lbpa-figure1-modal" class="journal-protocol"> Figure 1 </a></p> <!-- under data-target refer to a modal specifically made for each image under image modals section--> | <p><b>Results:</b><a data-toggle="modal" data-target="#lbpa-figure1-modal" class="journal-protocol"> Figure 1 </a></p> <!-- under data-target refer to a modal specifically made for each image under image modals section--> | ||
<p><b>Next Steps:</b> whatever comes next</p> | <p><b>Next Steps:</b> whatever comes next</p> | ||
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Revision as of 18:26, 6 August 2015
LABJOURNAL
BioSpray
Our forensic toolkit aims to use synthetic biology approaches to improve on the current methods used by crime scene investigators.
Summary
This week was all about getting started with the newly arrived biobrick BBa_K1058008
Aim of experiment:To prepare BBa_K1058008 for sequence confirmation, storage, and further processing.
Protocols Used:
Click here to see our plating protocol.
Click here to see our overnight culture protocol.
Results: Figure 1
Next Steps: whatever comes next
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment:Storage of BBa_K1058008
Protocols Used:
The agar stab received from the registry was transferred to an agar plate supplemented with 2x Chloramphenicol. The plate was incubated at 37°C for 16h and then stored at 4°C. Furthermore, an overnight culture was set up by inoculating 5ml LB, supplemented with chloramphenicol, with the agar stab. The overnight culture was incubated for 16h.
Aim of experiment:Purification and sequencing of BBa_K1058008
Protocols Used:
The overnight culture was retrieved and centrifuged at 4000rpm for 10min. The plasmid, pSB1C3, containing BBa_K1058008 was purified according to the miniprep protocol. The concentration was determined by nanodrop and found to be 190.9ng/µl. 2 Samples were submitted for sequencing, each containing 550ng DNA, 2µl of 3mM primer, and water filled up to a total volume of 30µl
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Week summary
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Aim of experiment: ultricies nec tellus eu
Protocols Used:
faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl. Morbi felis lectus, ultricies nec tellus eu, faucibus eleifend lectus. In molestie augue leo, id imperdiet ante imperdiet at. Quisque ultrices neque sit amet felis tincidunt tristique at sed nisi. Class aptent taciti sociosqu ad litora torquent per conubia nostra, per inceptos himenaeos. Ut suscipit ut massa in ultricies. Nunc nec tincidunt nunc, ut consectetur lorem. Sed non justo nislo nislo nisl.
Restriction digests were set up according to the table below. The appropriate reaction and restriction enzymes that were used is stated in the Journal section.
|
Post Plasmid Purification |
Post PCR |
gBlock |
|
|
DNA |
1mg |
50µl |
100ng |
|
Cutsmart Buffer (µl) |
6 |
6 |
6 |
|
Water (µl) |
As required |
1 |
As required |
|
Enzyme 1 (µl) |
1.5 |
1.5 |
1.5 |
|
Enzyme 2 (µl) |
1.5 |
1.5 |
1.5 |
|
Total Volume (µl) |
30 |
60 |
30 |
The digests were incubated at 37oC for 3 hours.
Note that when performing plasmid digestions, 2.5µl of alkaline phosphatase and 2.5µl of its buffer was added at the 2 hour and 2.5 hour mark.
Ligation reactions were set up according to the table below. The vector to insert ratio is stated for each specific reaction in the Journal section.
|
Vector (µl) |
2:1 (Vector: Insert) [µl] |
3:1 (Vector: Insert) [µl] |
4:1 (Vector: Insert) [µl] |
|
|
T4 DNA Ligase |
1 |
1 |
1 |
1 |
|
Ligase Buffer |
1 |
1 |
1 |
1 |
|
Vector |
1 |
1 |
1 |
1 |
|
Insert |
- |
2 |
3 |
4 |
|
H20 |
7 |
5 |
4 |
3 |
The reactions were incubated at room temperature for at least 3 hours.
'