Team:USC/Notebook/Week13

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<h1 style="font-family:Verdana;font-weight:700;">Laboratory Notebook</h1>
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<td style="text-align:center; font-family:Verdana; font-weight:700;">
[[Team:USC/Notebook/Brainstorming|Brainstorming]]
[[Team:USC/Notebook/Brainstorming|Brainstorming]]
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[[File:Brain_storm.JPG]]
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[[File:Brain_storm.JPG|link=Team:USC/Notebook/Brainstorming|Brainstorming]]
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[[Team:USC/Notebook/Brainstorming|Week 1]]
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[[Team:USC/Notebook/Week1|Week 1]]
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[[File:week1.jpg]]
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[[File:week1.jpg|link=Team:USC/Notebook/Week1|Week 1]]
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[[Team:USC/Notebook/Week2|Week 2]]
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[[File:week2.jpg|link=Team:USC/Notebook/Week2|Week 2]]
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[[Team:USC/Notebook/Brainstorming|Week 2]]
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[[Team:USC/Notebook/Week3|Week 3]]
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[[File:week2.jpg]]
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[[File:week3.jpg|link=Team:USC/Notebook/Week3|Week 3]]
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[[Team:USC/Notebook/Brainstorming|Week 3]]
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[[Team:USC/Notebook/Week4|Week 4]]
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[[File:week4.jpg|link=Team:USC/Notebook/Week4|Week 4]]
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[[Team:USC/Notebook/Week5|Week 5]]
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[[File:week5.jpg|link=Team:USC/Notebook/Week5|Week 5]]
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[[Team:USC/Notebook/Week6|Week 6]]
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[[File:week6.jpg|link=Team:USC/Notebook/Week6|Week 6]]
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[[Team:USC/Notebook/Week7|Week 7]]
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[[File:week7.jpg|link=Team:USC/Notebook/Week7|Week 7]]
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[[Team:USC/Notebook/Brainstorming|Week 7]]
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[[Team:USC/Notebook/Week8|Week 8]]
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[[File:week7.jpg]]
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[[File:week8.jpg|link=Team:USC/Notebook/Week8|Week 8]]
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[[Team:USC/Notebook/Week9|Week 9]]
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[[File:week9.jpg|link=Team:USC/Notebook/Week9|Week 9]]
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[[Team:USC/Notebook/Week10|Week 10]]
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[[File:week10.jpg|link=Team:USC/Notebook/Week10|Week 10]]
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[[Team:USC/Notebook/Week11|Week 11]]
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[[File:week10.jpg]]
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[[File:week11.jpg|link=Team:USC/Notebook/Week11|Week 11]]
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[[Team:USC/Notebook/Brainstorming|Week 11]]
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[[Team:USC/Notebook/Week12|Week 12]]
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[[File:week11.jpg]]
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[[File:week12.jpg|link=Team:USC/Notebook/Week12|Week 12]]
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[[Team:USC/Notebook/Brainstorming|Week 12]]
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[[Team:USC/Notebook/Week13|Week 13]]
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[[File:week12.jpg]]
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[[File:week13.jpg|link=Team:USC/Notebook/Week13|Week 13]]
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[[Team:USC/Notebook/Brainstorming|Week 13]]
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[[Team:USC/Notebook/Week14|Week 14]]
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[[File:week14.jpg|link=Team:USC/Notebook/Week14|Week 14]]
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<h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 1:'''</h3>
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<span style="font-family:Verdana; font-weight:700;"> 06/07/2011 </span>
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<h3 style="font-family:Verdana; font-weight:700;background-color: #F0F0F0;">'''Week 13:'''</h3>
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<span style="font-family:Verdana; font-weight:700;"> 09/06/2011 </span>
<br />
<br />
-
Transformation plasmids into DH5α Competent cells
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1. Digest/ ligate casO+PCDF
<br />
<br />
-
Plasmids are from iGEM plates:
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2. Transform ligated products into BL21 with tetO::GFP
<br />
<br />
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A. Plate 2 Well 7E  (AHL signaling)
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3. Transform CRISPR GFP#7 into BL21 with tetO::GFP
<br />
<br />
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B. Plate 3 Well 20F  (LovTAP composite)
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4. Inoculate HT5 with/without IPTG
<br />
<br />
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C. Plate 4 Well 16O  (LovTAP Composite)
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5. Inoculate HT115+tetO::GFP
<br />
<br />
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D. Plate 1 Well 2B  (GFP+PEST191)
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6. Purify casO (PCR products)
<br />
<br />
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E. Plate 2 Well 4A  (Yeast ADH1 promoter)
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7. Measure the DNA of casO::PCDF
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<span style="font-family:Verdana; font-weight:700;"> 06/08/2011 </span>
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<span style="font-family:Verdana; font-weight:700;"> 09/07/2011 </span>
<br />
<br />
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Transformation of E. Coli, Plasmids are from iGEM plates:
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1. Make competent cells as follows:
<br />
<br />
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Plasmids are from iGEM plates:
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BL21 (tetO::GFP, CRISPR::GFP)
<br />
<br />
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A. Plate 2 Well 13J
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HT115(tetO::GFP, CRISPR::GFP)
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B. Plate 1 Well 20F
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HT115(tetO::GFP)
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C. Plate 4 Well 16M
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2. Take OD reading of HT115 with/ without IPTG
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D. Plate 1 Well 16K
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3. Digest/ligate casO+PCDF
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<br />
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E. Plate 3 Well 14H
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4. Transform CRISPR#7 into tetO cells(BL21)
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F. Plate 4 Well 6O
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5. Take equal amount of E.Coli, miniprep, transform into new DH5α, select on amp.
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<br />
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<span style="font-family:Verdana; font-weight:700;"> 06/09/2011 </span>
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<span style="font-family:Verdana; font-weight:700;"> 09/19/2011 </span>
<br />
<br />
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1. Inoculation for all the plasmid we have transformed before.
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1. Growth test, grow all the bacteria overnight<br />
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Observation: all transformation except BBa-I15010
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<br />
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2. PCR cas3+casO from gDNA
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<br />
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<span style="font-family:Verdana; font-weight:700;"> 06/10/2011 </span>
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<span style="font-family:Verdana; font-weight:700;"> 09/08/2011 </span>
<br />
<br />
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1. Freeze the inoculated culture
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1.     OD reading
<br />
<br />
-
2. Observation: BBa-I15008 and BBa-I63009don’t have as much growth as others
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2. Digest/ligate reaction
 +
<br />
 +
tetR with pSB1k3
 +
<br />
 +
tetO with pSB1k3
 +
<br />
 +
casO with PCDF
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<br />
 +
Transform into DH5α, and also IPTG samples into DH5α
<br />
<br />
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<span style="font-family:Verdana; font-weight:700;"> 06/11/2011 </span>
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<span style="font-family:Verdana; font-weight:700;"> 09/09/2011 </span>
<br />
<br />
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1. Purify plasmid DNA from the culture
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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 />
<br />
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</table>
 

Latest revision as of 03:58, 29 September 2011

USC Banner.jpg


Laboratory Notebook

Brainstorming
Brainstorming

Week 1
Week 1

Week 2
Week 2

Week 3
Week 3

Week 4
Week 4

Week 5
Week 5

Week 6
Week 6

Week 7
Week 7

Week 8
Week 8

Week 9
Week 9

Week 10
Week 10

Week 11
Week 11

Week 12
Week 12

Week 13
Week 13

Week 14
Week 14

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

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