"
Team:Frankfurt/Notebook/Labwork
From 2013.igem.org
| Line 21: | Line 21: | ||
Expectation: 4 Bands with 2246, 3196, 2344, 3624 bp. Identification of one positive clone with the EcoRI-HF digest. | Expectation: 4 Bands with 2246, 3196, 2344, 3624 bp. Identification of one positive clone with the EcoRI-HF digest. | ||
<br> | <br> | ||
| - | 2. Production of competent DH5alpha | + | 2. Production of competent DH5alpha <br> |
| - | 3. Digest of plasmid p426 (mevalonate) with and without | + | 3. Digest of plasmid p426 (mevalonate) with and without insert. <br> |
| - | 4. Production of Cryo-Stocks of the p426 plasmids. | + | 4. Production of Cryo-Stocks of the p426 plasmids. <br> |
| - | 5. Transformation with the positive clone | + | 5. Transformation with the positive clone <br> |
| - | 6. Overnight cultures of yeast with the mevalonate plasmid. | + | 6. Overnight cultures of yeast with the mevalonate plasmid. <br> |
| - | 7. Making cultures of p426 with and without insert for GC usage. | + | 7. Making cultures of p426 with and without insert for GC usage. <br> |
| - | * Cultures were incubated for around 48 hours and harvested at an OD600 of 2.3 | + | * Cultures were incubated for around 48 hours and harvested at an OD600 of 2.3 <br> |
== September 2013 == __NOEDITSECTION__ | == September 2013 == __NOEDITSECTION__ | ||
{{:Team:Frankfurt/Footer}} | {{:Team:Frankfurt/Footer}} | ||
Revision as of 21:22, 4 October 2013
Contents |
Labwork
Unfortunately making arrangements for the lab this year and applying for funding consumed more time than expected resulting in less time for the actual project.
July 2013
1. Arrangements for labwork such as preparing material
- see protocols for the production of competent (E.coli, S.cerevisiae) and corresponding media (LB, YEPD, SCD-ura,…)
2. Purchase/Organization of the equipment (reaction tubes, glass bottles, pipette tips,..)
August 2013
Recapitulation of the work done in 2012 : restriction digest of plasmids to make sure we can use them:
1. Digests with enzymes: BsaAI, EcoRI-HF, XbaI
example:
- 1 µl DNA (~500nm)
- 0.5 µl BsaAI (5 units / µl -> one unit cuts once within one hour)
- 1 µl Buffer 4 (10x)
- 7,5 µl ddH2O
Expectation: 4 Bands with 2246, 3196, 2344, 3624 bp. Identification of one positive clone with the EcoRI-HF digest.
2. Production of competent DH5alpha
3. Digest of plasmid p426 (mevalonate) with and without insert.
4. Production of Cryo-Stocks of the p426 plasmids.
5. Transformation with the positive clone
6. Overnight cultures of yeast with the mevalonate plasmid.
7. Making cultures of p426 with and without insert for GC usage.
- Cultures were incubated for around 48 hours and harvested at an OD600 of 2.3
September 2013
