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Team:USC/Notebook/Week6
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<table> | <table> | ||
<tr> | <tr> | ||
| - | <h1 style="font-family:Verdana;font-weight:700;"> | + | <h1 style="font-family:Verdana;font-weight:700;">Laboratory Notebook</h1> |
</tr> | </tr> | ||
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[[Team:USC/Notebook/Brainstorming|Brainstorming]] | [[Team:USC/Notebook/Brainstorming|Brainstorming]] | ||
<br/> | <br/> | ||
| - | [[File:Brain_storm.JPG]] | + | [[File:Brain_storm.JPG|link=Team:USC/Notebook/Brainstorming|Brainstorming]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week1|Week 1]] | [[Team:USC/Notebook/Week1|Week 1]] | ||
<br/> | <br/> | ||
| - | [[File:week1.jpg]] | + | [[File:week1.jpg|link=Team:USC/Notebook/Week1|Week 1]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week2|Week 2]] | [[Team:USC/Notebook/Week2|Week 2]] | ||
<br/> | <br/> | ||
| - | [[File:week2.jpg]] | + | [[File:week2.jpg|link=Team:USC/Notebook/Week2|Week 2]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week3|Week 3]] | [[Team:USC/Notebook/Week3|Week 3]] | ||
<br/> | <br/> | ||
| - | [[File:week3.jpg]] | + | [[File:week3.jpg|link=Team:USC/Notebook/Week3|Week 3]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week4|Week 4]] | [[Team:USC/Notebook/Week4|Week 4]] | ||
<br/> | <br/> | ||
| - | [[File:week4.jpg]] | + | [[File:week4.jpg|link=Team:USC/Notebook/Week4|Week 4]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week5|Week 5]] | [[Team:USC/Notebook/Week5|Week 5]] | ||
<br/> | <br/> | ||
| - | [[File:week5.jpg]] | + | [[File:week5.jpg|link=Team:USC/Notebook/Week5|Week 5]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week6|Week 6]] | [[Team:USC/Notebook/Week6|Week 6]] | ||
<br/> | <br/> | ||
| - | [[File:week6.jpg]] | + | [[File:week6.jpg|link=Team:USC/Notebook/Week6|Week 6]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week7|Week 7]] | [[Team:USC/Notebook/Week7|Week 7]] | ||
<br/> | <br/> | ||
| - | [[File:week7.jpg]] | + | [[File:week7.jpg|link=Team:USC/Notebook/Week7|Week 7]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week8|Week 8]] | [[Team:USC/Notebook/Week8|Week 8]] | ||
<br/> | <br/> | ||
| - | [[File:week8.jpg]] | + | [[File:week8.jpg|link=Team:USC/Notebook/Week8|Week 8]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week9|Week 9]] | [[Team:USC/Notebook/Week9|Week 9]] | ||
<br/> | <br/> | ||
| - | [[File:week9.jpg]] | + | [[File:week9.jpg|link=Team:USC/Notebook/Week9|Week 9]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week10|Week 10]] | [[Team:USC/Notebook/Week10|Week 10]] | ||
<br/> | <br/> | ||
| - | [[File:week10.jpg]] | + | [[File:week10.jpg|link=Team:USC/Notebook/Week10|Week 10]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week11|Week 11]] | [[Team:USC/Notebook/Week11|Week 11]] | ||
<br/> | <br/> | ||
| - | [[File:week11.jpg]] | + | [[File:week11.jpg|link=Team:USC/Notebook/Week11|Week 11]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week12|Week 12]] | [[Team:USC/Notebook/Week12|Week 12]] | ||
<br/> | <br/> | ||
| - | [[File:week12.jpg]] | + | [[File:week12.jpg|link=Team:USC/Notebook/Week12|Week 12]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week13|Week 13]] | [[Team:USC/Notebook/Week13|Week 13]] | ||
<br/> | <br/> | ||
| - | [[File:week13.jpg]] | + | [[File:week13.jpg|link=Team:USC/Notebook/Week13|Week 13]] |
</td> | </td> | ||
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[[Team:USC/Notebook/Week14|Week 14]] | [[Team:USC/Notebook/Week14|Week 14]] | ||
<br/> | <br/> | ||
| - | [[File:week14.jpg]] | + | [[File:week14.jpg|link=Team:USC/Notebook/Week14|Week 14]] |
</td> | </td> | ||
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</table> | </table> | ||
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<h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 6:'''</h3> | <h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 6:'''</h3> | ||
Latest revision as of 03:57, 29 September 2011
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Week 6:
07/11/2011
1. inoculate prs 423+MET25, prs 424+MET25, prs 424+MET25
2. PCR TPS1, MSN2, ELO1, OLE1
3. Inoculate and mini-prep:
PCDF(s) and PCOLA(K)
07/12/2011
1. Observations: PCR products of CASO didn’t work
2. Test:
Old forward primer with new reverse primer
New forward primer with old reverse primer
3. Purify ligated vectors:
prs 423+MET25, prs 424+MET25, prs 424+MET25
4. Run a gel for PCR genes
5. Digest ligated vectors and run a gel to test them
Observations: ligated products didn’t work, all genes work
07/13/2011
1. Make competent cells BL21
2. Transformation and inoculation :
3. 1124, 1126, 1125, 1128, 1129, 1130, PCDF(5 µL step) and PCDF(10 µL step)
4. Run a gel for CAS3 and PCDF
5. Purify DNA:
PCDF, CRISPR, pSB1C3, tet spacer, GFP spacer, CAS3 #1 and CAS3 #2
6. PCR CASO with E, E1, E2 and AP1 and AP2 (primers)
07/14/2011
1. Make competent cells , check the OD after incubating in shaker until OD~0.25
Check every 30 mins
2. Inoculate and transform PCOLA
3. Transform CRISPR
4. Mini-prep 1124-1130, PCDF
5. Gel verify CASO by PCR
6. Inoculate DH5α and BL21
7. Digest tetR
07/15/2011
1. Transform pSB1C3 tetR, CRISPR tetR, CRISPR GFP, CRISPR
2. Run a gel to test:
Ladder, CAS3 #2, CAS3 #3, PCDF, CRISPR, tetR spacer, GFP spacer, pSB1C3
</table>
