Team:USC/Notebook/Week13

From 2011.igem.org

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<h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 13:'''</h3>
<h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 13:'''</h3>
 +
 +
<span style="font-family:Verdana; font-weight:700;"> 09/06/2011 </span>
 +
<br />
 +
1. Digest/ ligate casO+PCDF
 +
<br />
 +
2. Transform ligated products into BL21 with tetO::GFP
 +
<br />
 +
3. Transform CRISPR GFP#7 into BL21 with tetO::GFP
 +
<br />
 +
4. Inoculate HT5 with/without IPTG
 +
<br />
 +
5. Inoculate HT115+tetO::GFP
 +
<br />
 +
6. Purify casO (PCR products)
 +
<br />
 +
7. Measure the DNA of casO::PCDF
 +
 +
 +
<span style="font-family:Verdana; font-weight:700;"> 09/07/2011 </span>
 +
<br />
 +
1. Make competent cells as follows:
 +
<br />
 +
BL21 (tetO::GFP, CRISPR::GFP)
 +
<br />
 +
HT115(tetO::GFP, CRISPR::GFP)
 +
<br />
 +
HT115(tetO::GFP)
 +
<br />
 +
2. Take OD reading of HT115 with/ without IPTG
 +
<br />
 +
3. Digest/ligate casO+PCDF
 +
<br />
 +
4. Transform CRISPR#7 into tetO cells(BL21)
 +
<br />
 +
5. Take equal amount of E.Coli, miniprep, transform into new DH5α, select on amp.
 +
<br />
 +
 +
 +
<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;"> 09/08/2011 </span>
 +
<br />
 +
1.      OD reading
 +
<br />
 +
2. Digest/ligate reaction
 +
<br />
 +
tetR with pSB1k3
 +
<br />
 +
tetO with pSB1k3
 +
<br />
 +
casO with PCDF
 +
<br />
 +
Transform into DH5α, and also IPTG samples into DH5α
 +
<br />
 +
 +
<span style="font-family:Verdana; font-weight:700;"> 09/09/2011 </span>
 +
<br />
 +
1.      Make BL21(tetO, CRISPR-GFP #7) competent and make freeze stock
 +
<br />
 +
2. Make HT115(tetO, CRISPR-GFP #7) competent and make freeze stock
 +
<br />
 +
3. Make HT115(CRISPR-GFP #7) competent and make freeze stock
 +
<br />
 +
 +
 +
<tr>
 +
<td colspan="7">
 +
<h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 14:'''</h3>
<span style="font-family:Verdana; font-weight:700;"> 09/13/2011 </span>
<span style="font-family:Verdana; font-weight:700;"> 09/13/2011 </span>

Revision as of 06:09, 25 September 2011

USC-logo.jpg IGEM2011-logo.jpg IGEM-logo.jpg
USC Banner.jpg

Brainstorming
Brain storm.JPG

Week 1
Week1.jpg

Week 2
Week2.jpg

Week 3
Week3.jpg

Week 4
Week4.jpg

Week 5
Week5.jpg

Week 6
Week6.jpg

Week 7
Week7.jpg

Week 8
Week8.jpg

Week 9
Week9.jpg

Week 10
Week10.jpg

Week 11
Week11.jpg

Week 12
Week12.jpg

Week 13
Week13.jpg

Week 13:

09/06/2011
1. Digest/ ligate casO+PCDF
2. Transform ligated products into BL21 with tetO::GFP
3. Transform CRISPR GFP#7 into BL21 with tetO::GFP
4. Inoculate HT5 with/without IPTG
5. Inoculate HT115+tetO::GFP
6. Purify casO (PCR products)
7. Measure the DNA of casO::PCDF


09/07/2011
1. Make competent cells as follows:
BL21 (tetO::GFP, CRISPR::GFP)
HT115(tetO::GFP, CRISPR::GFP)
HT115(tetO::GFP)
2. Take OD reading of HT115 with/ without IPTG
3. Digest/ligate casO+PCDF
4. Transform CRISPR#7 into tetO cells(BL21)
5. Take equal amount of E.Coli, miniprep, transform into new DH5α, select on amp.


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

2. PCR cas3+casO from gDNA


09/08/2011
1. OD reading
2. Digest/ligate reaction
tetR with pSB1k3
tetO with pSB1k3
casO with PCDF
Transform into DH5α, and also IPTG samples into DH5α

09/09/2011
1. Make BL21(tetO, CRISPR-GFP #7) competent and make freeze stock
2. Make HT115(tetO, CRISPR-GFP #7) competent and make freeze stock
3. Make HT115(CRISPR-GFP #7) competent and make freeze stock


Week 14:

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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