Team:USC/Notebook/Week13

From 2011.igem.org

(Difference between revisions)
Line 160: Line 160:
1. Inoculate the following:
1. Inoculate the following:
<br />
<br />
-
    BL21 tetO::GFP
+
BL21 tetO::GFP
<br />
<br />
-
2. BL21 tetO::GFP, CRISPR::GFP#7
+
BL21 tetO::GFP, CRISPR::GFP#7
-
3. BL21 tetO::GFP, CRISPR::GFP#7, cas3
+
-
4. HT115 tetO::GFP
+
-
5. HT115 tetO::GFP, CRISPR::GFP#7
+
-
6. HT115 tetO::GFP, CRISPR::GFP#7, cas3
+
-
7. HT115 CRISPR::GFP #7
+
<br />
<br />
-
C. Plate 4 Well 16O  (LovTAP Composite)
+
BL21 tetO::GFP, CRISPR::GFP#7, cas3
<br />
<br />
-
D. Plate 1 Well 2B  (GFP+PEST191)
+
HT115 tetO::GFP
<br />
<br />
-
E. Plate 2 Well 4A  (Yeast ADH1 promoter)
+
HT115 tetO::GFP, CRISPR::GFP#7
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7, cas3
 +
<br />
 +
HT115 CRISPR::GFP #7
 +
<span style="font-family:Verdana; font-weight:700;"> 09/19/2011 </span>
 +
<br />
 +
1. Growth test, grow all the bacteria overnight<br />
 +
<br />
 +
2. PCR cas3+casO from gDNA
 +
<br />
-
<span style="font-family:Verdana; font-weight:700;"> 06/10/2011 </span>
+
<span style="font-family:Verdana; font-weight:700;"> 09/20/2011 </span>
<br />
<br />
-
1. Freeze the inoculated culture
+
1.     Make biobrick of cas3 and casO
<br />
<br />
-
2. Observation: BBa-I15008 and BBa-I63009don’t have as much growth as others
+
2. Run a gel for cas3 and casO
 +
<br />
 +
3. Purify cas3 and casO
 +
<br />
 +
4. New experiment: dilution the bacteria to 1000 and 5000 cells and plate them
<br />
<br />
-
<span style="font-family:Verdana; font-weight:700;"> 06/11/2011 </span>
+
<span style="font-family:Verdana; font-weight:700;"> 09/21/2011 </span>
<br />
<br />
-
1. Purify plasmid DNA from the culture
+
1.     Do growth test for the following:
<br />
<br />
 +
BL21 tetO::GFP-IPTG
 +
<br />
 +
BL21 tetO::GFP+IPTG
 +
<br />
 +
BL21 tetO::GFP, CRISPR::GFP#7-IPTG
 +
<br />
 +
BL21 tetO::GFP, CRISPR::GFP#7+IPTG
 +
<br />
 +
BL21 tetO::GFP, CRISPR::GFP#7, cas3-IPTG
 +
<br />
 +
BL21 tetO::GFP, CRISPR::GFP#7, cas3+IPTG
 +
<br />
 +
BL21 tetO::GFP, CRISPR::GFP#7, cas3+IPTG an hour later
 +
<br />
 +
HT115 tetO::GFP-IPTG
 +
<br />
 +
HT115 tetO::GFP+IPTG
 +
<br />
 +
HT115 tetO::GFP+IPTG an hour later
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7-IPTG
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7+IPTG
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7+IPTG an hour later
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7, cas3-IPTG
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7, cas3+IPTG
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7, cas3+IPTG an hour later
 +
<br />
 +
HT115 CRISPR::GFP #7-IPTG
 +
<br />
 +
HT115 CRISPR::GFP #7+IPTG
 +
<br />
 +
HT115 CRISPR::GFP #7+IPTG an hour  later
 +
<br />
 +
BL21 tetO::GFP, CRISPR::GFP#7, cas3
 +
<br />
 +
HT115 tetO::GFP
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7
 +
<br />
 +
HT115 tetO::GFP, CRISPR::GFP#7, cas3
 +
<br />
 +
HT115 CRISPR::GFP #7
 +
<br />
 +
 +
</table>
</table>

Revision as of 05:35, 25 September 2011

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Brainstorming
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Week 1
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Week 2
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Week 3
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Week 4
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Week 5
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Week 6
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Week 7
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Week 8
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Week 9
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Week 10
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Week 11
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Week 12
Week12.jpg

Week 13
Week13.jpg

Week 13:

09/13/2011
1. Check quality of gDNA
2. Make HT115 + BL21 (tetO, CRISPR-GFP #7) competent cells, freeze stock
3. Miniprep PCDF+casO
C. Plate 4 Well 16O (LovTAP Composite)
4. Digest tetO/tetR

09/15/2011
1. Run a gel of all minipreped PCDF+casO

2. PCR HT115, DH5α2, DH5α3 with primers

09/18/2011
1. Inoculate the following:
BL21 tetO::GFP
BL21 tetO::GFP, CRISPR::GFP#7
BL21 tetO::GFP, CRISPR::GFP#7, cas3
HT115 tetO::GFP
HT115 tetO::GFP, CRISPR::GFP#7
HT115 tetO::GFP, CRISPR::GFP#7, cas3
HT115 CRISPR::GFP #7


09/19/2011
1. Growth test, grow all the bacteria overnight

2. PCR cas3+casO from gDNA


09/20/2011
1. Make biobrick of cas3 and casO
2. Run a gel for cas3 and casO
3. Purify cas3 and casO
4. New experiment: dilution the bacteria to 1000 and 5000 cells and plate them

09/21/2011
1. Do growth test for the following:
BL21 tetO::GFP-IPTG
BL21 tetO::GFP+IPTG
BL21 tetO::GFP, CRISPR::GFP#7-IPTG
BL21 tetO::GFP, CRISPR::GFP#7+IPTG
BL21 tetO::GFP, CRISPR::GFP#7, cas3-IPTG
BL21 tetO::GFP, CRISPR::GFP#7, cas3+IPTG
BL21 tetO::GFP, CRISPR::GFP#7, cas3+IPTG an hour later
HT115 tetO::GFP-IPTG
HT115 tetO::GFP+IPTG
HT115 tetO::GFP+IPTG an hour later
HT115 tetO::GFP, CRISPR::GFP#7-IPTG
HT115 tetO::GFP, CRISPR::GFP#7+IPTG
HT115 tetO::GFP, CRISPR::GFP#7+IPTG an hour later
HT115 tetO::GFP, CRISPR::GFP#7, cas3-IPTG
HT115 tetO::GFP, CRISPR::GFP#7, cas3+IPTG
HT115 tetO::GFP, CRISPR::GFP#7, cas3+IPTG an hour later
HT115 CRISPR::GFP #7-IPTG
HT115 CRISPR::GFP #7+IPTG
HT115 CRISPR::GFP #7+IPTG an hour later
BL21 tetO::GFP, CRISPR::GFP#7, cas3
HT115 tetO::GFP
HT115 tetO::GFP, CRISPR::GFP#7
HT115 tetO::GFP, CRISPR::GFP#7, cas3
HT115 CRISPR::GFP #7


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