Team:Yale/Our Project/Notebook/Week 3
From 2010.igem.org
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<li>Began miniprep of overnight cultures, but in initial cell pelleting phase the centrifuge spun the tops of the tubes, spilling them. </li> | <li>Began miniprep of overnight cultures, but in initial cell pelleting phase the centrifuge spun the tops of the tubes, spilling them. </li> | ||
<li> Started over with the inoculation of two 5 mL liquid ampicillin LB cultures each of pSB74, R0011, C0012,J23114, and B0015 containing strains. </li> | <li> Started over with the inoculation of two 5 mL liquid ampicillin LB cultures each of pSB74, R0011, C0012,J23114, and B0015 containing strains. </li> | ||
- | <li> When OD | + | <li> When OD was approximately 0.6, took 0.5 mL of each solution and mixed with 0.5 mL of 50% filter-sterilized glycerol and set aside for glycerol stocks. </li> |
<li> Let cultures continue to grow all day, then miniprepped via the <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/miniprep"> microcentrifuge protocol, </a> eluting into 40 uL of water. </li> | <li> Let cultures continue to grow all day, then miniprepped via the <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/miniprep"> microcentrifuge protocol, </a> eluting into 40 uL of water. </li> | ||
</ul> | </ul> | ||
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<b>Nanodrop of 6/22 miniprep </b> <br/> | <b>Nanodrop of 6/22 miniprep </b> <br/> | ||
Nanodropped the ten minipreps from the day before and got the following results: | Nanodropped the ten minipreps from the day before and got the following results: | ||
+ | <table> | ||
+ | <tr> | ||
<td>Sample</td> | <td>Sample</td> | ||
<td>J23114a</td><td>J23114b</td><td>C0012a</td><td>C0012b</td><td>R0011a</td><td>R0011b</td><td>B0015a</td><td>B0015b</td><td>pSB74a</td><td>pSB74b</td> | <td>J23114a</td><td>J23114b</td><td>C0012a</td><td>C0012b</td><td>R0011a</td><td>R0011b</td><td>B0015a</td><td>B0015b</td><td>pSB74a</td><td>pSB74b</td> | ||
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So that can readily determine whether bacteria are producing hydrogen sulfide, created a batch of TSI agar slants according to the following <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/TSI_agar"> recipe </a>, with ampicillin as the added antibiotic and tubes of the medium allowed to solidify on an angle to produce a characteristic slant. If a culture containing thiosulfate reductase is grown in such a slant, it will turn the medium black by precipitating out black iron sulfide, an obvious visual cue of hydrogen sulfide production. <br/> | So that can readily determine whether bacteria are producing hydrogen sulfide, created a batch of TSI agar slants according to the following <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/TSI_agar"> recipe </a>, with ampicillin as the added antibiotic and tubes of the medium allowed to solidify on an angle to produce a characteristic slant. If a culture containing thiosulfate reductase is grown in such a slant, it will turn the medium black by precipitating out black iron sulfide, an obvious visual cue of hydrogen sulfide production. <br/> | ||
<b> Transformation of BBa_P0312</b> <br/> | <b> Transformation of BBa_P0312</b> <br/> | ||
- | After realizing that Biobrick P0312 can be used in place of C0012 and R0011, | + | After realizing that Biobrick P0312 can be used in place of C0012 and R0011, transformed BL21 with P0312 according to the <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/transformation"> standard transformation protocol </a> <br/> |
<i> This day's labwork is also recorded on pages 18, 20, and 21 of the hard copy lab notebook. </i> | <i> This day's labwork is also recorded on pages 18, 20, and 21 of the hard copy lab notebook. </i> | ||
<!------------- Wednesday -------------> | <!------------- Wednesday -------------> |
Revision as of 08:24, 27 October 2010
our project
lab notebook: week 3 (6/21-6/27)
- Monday 6/21--Miniprep Biobrick plasmids from overnight culture, but a mix up samples, necessitating a diagnostic digest. See more/less
- Tuesday 6/22--Miniprep redos See more/less
- Wednesday 6/23--Making TSI agar slants & transformation of P0312 See more/less
- Thursday See more/less
- Friday See more/less
- 6/21-AM miniprep but mixed up tubes--performed EcoR I/Pst I digest & gel to help distinguish tubes, though not all of them can be identified. Inoculated liquid cultures of transformants (again)
- 6/22 miniprep again