Virginia Commonwealth/14 July 2009

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Latest revision as of 20:45, 17 October 2009

May
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				[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/1_May_2009&action=edit 1]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/2_May_2009&action=edit 2]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/3_May_2009&action=edit 3]
[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_May_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_May_2009&action=edit 5]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/6_May_2009&action=edit 6]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/7_May_2009&action=edit 7]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/8_May_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/9_May_2009&action=edit 9]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/10_May_2009&action=edit 10]
[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_May_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_May_2009&action=edit 12]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/13_May_2009&action=edit 13]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/14_May_2009&action=edit 14]	[http://2009.igem.org/Virginia_Commonwealth/15_May_2009 15]	[http://2009.igem.org/Virginia_Commonwealth/16_May_2009 16]	[http://2009.igem.org/Virginia_Commonwealth/17_May_2009 17]
[http://2009.igem.org/Virginia_Commonwealth/18_May_2009 18]	[http://2009.igem.org/Virginia_Commonwealth/19_May_2009 19]	[http://2009.igem.org/Virginia_Commonwealth/20_May_2009 20]	[http://2009.igem.org/Virginia_Commonwealth/21_May_2009 21]	[http://2009.igem.org/Virginia_Commonwealth/22_May_2009 22]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/23_May_2009&action=edit 23]	[http://2009.igem.org/Virginia_Commonwealth/24_May_2009 24]
[http://2009.igem.org/Virginia_Commonwealth/25_May_2009 25]	[http://2009.igem.org/Virginia_Commonwealth/26_May_2009 26]	[http://2009.igem.org/Virginia_Commonwealth/27_May_2009 27]	[http://2009.igem.org/Virginia_Commonwealth/28_May_2009 28]	[http://2009.igem.org/Virginia_Commonwealth/29_May_2009 29]	[http://2009.igem.org/Virginia_Commonwealth/30_May_2009 30]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/31_May_2009&action=edit 31]

June
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[http://2009.igem.org/Virginia_Commonwealth/1_June_2009 1]	[http://2009.igem.org/Virginia_Commonwealth/2_June_2009 2]	[http://2009.igem.org/Virginia_Commonwealth/3_June_2009 3]	[http://2009.igem.org/Virginia_Commonwealth/4_June_2009 4]	[http://2009.igem.org/Virginia_Commonwealth/5_June_2009 5]	[http://2009.igem.org/Virginia_Commonwealth/6_June_2009 6]	[http://2009.igem.org/Virginia_Commonwealth/7_June_2009 7]
[http://2009.igem.org/Virginia_Commonwealth/8_June_2009 8]	[http://2009.igem.org/Virginia_Commonwealth/9_June_2009 9]	[http://2009.igem.org/Virginia_Commonwealth/10_June_2009 10]	[http://2009.igem.org/Virginia_Commonwealth/11_June_2009 11]	[http://2009.igem.org/Virginia_Commonwealth/12_June_2009 12]	[http://2009.igem.org/Virginia_Commonwealth/13_June_2009 13]	[http://2009.igem.org/Virginia_Commonwealth/14_June_2009 14]
[http://2009.igem.org/Virginia_Commonwealth/15_June_2009 15]	[http://2009.igem.org/Virginia_Commonwealth/16_June_2009 16]	[http://2009.igem.org/Virginia_Commonwealth/17_June_2009 17]	[http://2009.igem.org/Virginia_Commonwealth/18_June_2009 18]	[http://2009.igem.org/Virginia_Commonwealth/19_June_2009 19]	[http://2009.igem.org/Virginia_Commonwealth/20_June_2009 20]	[http://2009.igem.org/Virginia_Commonwealth/21_June_2009 21]
[http://2009.igem.org/Virginia_Commonwealth/22_June_2009 22]	[http://2009.igem.org/Virginia_Commonwealth/23_June_2009 23]	[http://2009.igem.org/Virginia_Commonwealth/24_June_2009 24]	[http://2009.igem.org/Virginia_Commonwealth/25_June_2009 25]	[http://2009.igem.org/Virginia_Commonwealth/26_June_2009 26]	[http://2009.igem.org/Virginia_Commonwealth/27_June_2009 27]	[http://2009.igem.org/Virginia_Commonwealth/28_June_2009 28]
[http://2009.igem.org/Virginia_Commonwealth/29_June_2009 29]	[http://2009.igem.org/Virginia_Commonwealth/30_June_2009 30]

July
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		[http://2009.igem.org/Virginia_Commonwealth/1_July_2009 1]	[http://2009.igem.org/Virginia_Commonwealth/2_July_2009 2]	[http://2009.igem.org/Virginia_Commonwealth/3_July_2009 3]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_July_2009&action=edit 4]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_July_2009&action=edit 5]
[http://2009.igem.org/Virginia_Commonwealth/6_July_2009 6]	[http://2009.igem.org/Virginia_Commonwealth/7_July_2009 7]	[http://2009.igem.org/Virginia_Commonwealth/8_July_2009 8]	[http://2009.igem.org/Virginia_Commonwealth/9_July_2009 9]	[http://2009.igem.org/Virginia_Commonwealth/10_July_2009 10]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_July_2009&action=edit 11]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_July_2009&action=edit 12]
[http://2009.igem.org/Virginia_Commonwealth/13_July_2009 13]	[http://2009.igem.org/Virginia_Commonwealth/14_July_2009 14]	[http://2009.igem.org/Virginia_Commonwealth/15_July_2009 15]	[http://2009.igem.org/Virginia_Commonwealth/16_July_2009 16]	[http://2009.igem.org/Virginia_Commonwealth/17_July_2009 17]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/18_July_2009&action=edit 18]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/19_July_2009&action=edit 19]
[http://2009.igem.org/Virginia_Commonwealth/20_July_2009 20]	[http://2009.igem.org/Virginia_Commonwealth/21_July_2009 21]	[http://2009.igem.org/Virginia_Commonwealth/22_July_2009 22]	[http://2009.igem.org/Virginia_Commonwealth/23_July_2009 23]	[http://2009.igem.org/Virginia_Commonwealth/24_July_2009 24]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/25_July_2009&action=edit 25]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/26_July_2009&action=edit 26]
[http://2009.igem.org/Virginia_Commonwealth/27_July_2009 27]	[http://2009.igem.org/Virginia_Commonwealth/28_July_2009 28]	[http://2009.igem.org/Virginia_Commonwealth/29_July_2009 29]	[http://2009.igem.org/Virginia_Commonwealth/30_July_2009 30]	[http://2009.igem.org/Virginia_Commonwealth/31_July_2009 31]

August
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					[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/1_August_2009&action=edit 1]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/2_August_2009&action=edit 2]
[http://2009.igem.org/Virginia_Commonwealth/3_August_2009 3]	[http://2009.igem.org/Virginia_Commonwealth/4_August_2009 4]	[http://2009.igem.org/Virginia_Commonwealth/5_August_2009 5]	[http://2009.igem.org/Virginia_Commonwealth/6_August_2009 6]	[http://2009.igem.org/Virginia_Commonwealth/7_August_2009 7]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/8_August_2009&action=edit 8]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/9_August_2009&action=edit 9]
[http://2009.igem.org/Virginia_Commonwealth/10_August_2009 10]	[http://2009.igem.org/Virginia_Commonwealth/11_August_2009 11]	[http://2009.igem.org/Virginia_Commonwealth/12_August_2009 12]	[http://2009.igem.org/Virginia_Commonwealth/13_August_2009 13]	[http://2009.igem.org/Virginia_Commonwealth/14_August_2009 14]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/15_August_2009&action=edit 15]	[http://2009.igem.org/Virginia_Commonwealth/16_August_2009 16]
[http://2009.igem.org/Virginia_Commonwealth/17_August_2009 17]	[http://2009.igem.org/Virginia_Commonwealth/18_August_2009 18]	[http://2009.igem.org/Virginia_Commonwealth/19_August_2009 19]	[http://2009.igem.org/Virginia_Commonwealth/20_August_2009 20]	[http://2009.igem.org/Virginia_Commonwealth/21_August_2009 21]	[http://2009.igem.org/Virginia_Commonwealth/22_August_2009 22]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/23_August_2009&action=edit 23]
[http://2009.igem.org/Virginia_Commonwealth/24_August_2009 24]	[http://2009.igem.org/Virginia_Commonwealth/25_August_2009 25]	[http://2009.igem.org/Virginia_Commonwealth/26_August_2009 26]	[http://2009.igem.org/Virginia_Commonwealth/27_August_2009 27]	[http://2009.igem.org/Virginia_Commonwealth/28_August_2009 28]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/29_August_2009&action=edit 29]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/30_August_2009&action=edit 30]
[http://2009.igem.org/Virginia_Commonwealth/31_August_2009 31]

September
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	[http://2009.igem.org/Virginia_Commonwealth/1_September_2009 1]	[http://2009.igem.org/Virginia_Commonwealth/2_September_2009 2]	[http://2009.igem.org/Virginia_Commonwealth/3_September_2009 3]	[http://2009.igem.org/Virginia_Commonwealth/4_September_2009 4]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_September_2009&action=edit 5]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/6_September_2009&action=edit 6]
[http://2009.igem.org/Virginia_Commonwealth/7_September_2009 7]	[http://2009.igem.org/Virginia_Commonwealth/8_September_2009 8]	[http://2009.igem.org/Virginia_Commonwealth/9_September_2009 9]	[http://2009.igem.org/Virginia_Commonwealth/10_September_2009 10]	[http://2009.igem.org/Virginia_Commonwealth/11_September_2009 11]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_September_2009&action=edit 12]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/13_September_2009&action=edit 13]
[http://2009.igem.org/Virginia_Commonwealth/14_September_2009 14]	[http://2009.igem.org/Virginia_Commonwealth/15_September_2009 15]	[http://2009.igem.org/Virginia_Commonwealth/16_September_2009 16]	[http://2009.igem.org/Virginia_Commonwealth/17_September_2009 17]	[http://2009.igem.org/Virginia_Commonwealth/18_September_2009 18]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/19_September_2009&action=edit 19]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/20_September_2009&action=edit 20]
[http://2009.igem.org/Virginia_Commonwealth/21_September_2009 21]	[http://2009.igem.org/Virginia_Commonwealth/22_September_2009 22]	[http://2009.igem.org/Virginia_Commonwealth/23_September_2009 23]	[http://2009.igem.org/Virginia_Commonwealth/24_September_2009 24]	[http://2009.igem.org/Virginia_Commonwealth/25_September_2009 25]	[http://2009.igem.org/Virginia_Commonwealth/26_September_2009 26]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/27_September_2009&action=edit 27]
[http://2009.igem.org/Virginia_Commonwealth/28_September_2009 28]	[http://2009.igem.org/Virginia_Commonwealth/29_September_2009 29]	[http://2009.igem.org/Virginia_Commonwealth/30_September_2009 30]

October
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			[http://2009.igem.org/Virginia_Commonwealth/1_October_2009 1]	[http://2009.igem.org/Virginia_Commonwealth/2_October_2009 2]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/3_October_2009&action=edit 3]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_October_2009&action=edit 4]
[http://2009.igem.org/Virginia_Commonwealth/5_October_2009 5]	[http://2009.igem.org/Virginia_Commonwealth/6_October_2009 6]	[http://2009.igem.org/Virginia_Commonwealth/7_October_2009 7]	[http://2009.igem.org/Virginia_Commonwealth/8_October_2009 8]	[http://2009.igem.org/Virginia_Commonwealth/9_October_2009 9]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/10_October_2009&action=edit 10]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_October_2009&action=edit 11]
[http://2009.igem.org/Virginia_Commonwealth/12_October_2009 12]	[http://2009.igem.org/Virginia_Commonwealth/13_October_2009 13]	[http://2009.igem.org/Virginia_Commonwealth/14_October_2009 14]	[http://2009.igem.org/Virginia_Commonwealth/15_October_2009 15]	[http://2009.igem.org/Virginia_Commonwealth/16_October_2009 16]	[http://2009.igem.org/Virginia_Commonwealth/17_October_2009 17]	[http://2009.igem.org/Virginia_Commonwealth/18_October_2009 18]
[http://2009.igem.org/Virginia_Commonwealth/19_October_2009 19]	[http://2009.igem.org/Virginia_Commonwealth/20_October_2009 20]	[http://2009.igem.org/Virginia_Commonwealth/21_October_2009 21]	[http://2009.igem.org/Virginia_Commonwealth/22_October_2009 22]	[http://2009.igem.org/Virginia_Commonwealth/23_October_2009 23]	[http://2009.igem.org/Virginia_Commonwealth/24_October_2009 24]	[http://2009.igem.org/Virginia_Commonwealth/25_October_2009 25]
[http://2009.igem.org/Virginia_Commonwealth/26_October_2009 26]	[http://2009.igem.org/Virginia_Commonwealth/27_October_2009 27]	[http://2009.igem.org/Virginia_Commonwealth/28_October_2009 28]	[http://2009.igem.org/Virginia_Commonwealth/29_October_2009 29]	[http://2009.igem.org/Virginia_Commonwealth/30_October_2009 30]	[http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/31_October_2009&action=edit 31]

Tuesday 14 July 2009

Results

Maria and Afton

All of the promoters showed up on the gel electrophoresis in a mostly straight line.
Transformation Results


Name	Plate	Growth Observation
Control (+) NEB10β	LB	lawn
Control (+) NEB10β shocked	LB	lawn
Control (-) NEB10β	Cm	no growth
J23106, J06702, pSB4C5 ligation	Cm	~30 small colonies

Miniprep Results:


Part No.	A260 (nm)	A280 (nm)	A260/A280	Vol. (µL)	Conc. (µg/mL)	Amt (µg)
J23101	0.050	0.039	1.20	47	75	4.70
J23102	0.030	0.025	1.20	58	45	3.48
J23103	0.044	0.032	1.38	56	66	4.93
J23104	0.033	0.028	1.18	54	49	3.56
J23105	0.037	0.029	1.28	32	56	1.78
J23107	0.054	0.040	1.35	47	81	3.81

Trentay 13:44, 14 July 2009 (UTC)

Gel Electrophoresis Results

from left to right: ladder, 100-105,107

Kevin and Adam

Miniprep results:


Part No.	A260 (nm)	A280 (nm)	A260/A280	Vol. (µL)	Conc. (µg/mL)	Amt (µg)
E0240	.033	.019	1.737	124	66	8.184
pSB4C5	.054	.036	1.5	123.5	108	13.34

Bussingkm 15:21, 14 July 2009 (UTC)

Craig and Clay

The ampicillin plates left overnight again displayed no growth. The electrotransformed parts will be re-plated onto ampicillin-selective plates as well as non-selective plates.

Tasks

Maria and Afton

Digest promoters: 101, 102, 103, 104
Possible Ligation of promoters with parts J06702, pSB4C5

Trentay 15:13, 14 July 2009 (UTC)

Kevin and Adam

perform complete digestion and ligation
construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106)

Bussingkm 15:20, 14 July 2009 (UTC)

Craig and Clay

The electrotransformed parts will be re-plated on ampicillin plates as well as on non-selective plates. Growth on non-selective plates will prove the existence of viable cells.

Wetlab

Maria and Afton

Spectophotometry of promoters: 101, 102, 103, 104, 105, 107
Digestion/Ligation of promoters
Electrophoresis of promoter digestions: 101, 102, 103, 104, 105, 107, (100)
Electrotransformation of ligated parts
- two Cm plate had an unidentified growth in the refrigerator.
- however, negative controls from the same plate stock show antibiotic is working
Picked colonies for overnight culture of J23106 w/ J06702 reporter
Made 3 overnight 5mL cultures of NEB 10 beta cell stock to prepare electrocompetent cells

Trentay 21:01, 14 July 2009 (UTC)

Kevin and Adam

pSB4C5, BBa_E0240, and BBa_J06702 were frozen in box C-11 of the glycerol stocks

Bussingkm 15:22, 14 July 2009 (UTC)

Craig and Clay

The electrotransformed parts were again plated on ampicillin-selective plates. They were also plated on non-selective media. One hundred microliters was used for each plate rather than the standard 35 microliters.

@@ Line 40: / Line 40: @@
 ===Results===
 ''Maria and Afton''
-* (+) Control: NEB 10 beta on LB
-**lawn growth
-* (+) Control: NEB 10 beta on LB shocked
-**lawn growth
-* (-) Control: NEB 10 beta on Cm
-**no growth
-* NEB 10 beta with BioBrick part (J23106, J06702, pSB4C5)
-**Approximately 30 small colonies
 * All of the promoters showed up on the gel electrophoresis in a mostly straight line.
+* Transformation Results
+{| cellpadding="5" border="1" padding="5"
+|+
+|-
+! Name !! Plate !! Growth Observation
+|-
+| Control (+) NEB10β || LB || lawn
+|-
+| Control (+) NEB10β shocked || LB || lawn
+|-
+| Control (-) NEB10β || Cm || no growth
+|-
+| J23106, J06702, pSB4C5 ligation || Cm || ~30 small colonies
+|}
+* Miniprep Results:
+{| cellpadding="5" border="1" padding="5"
+|+
+|-
+! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !!  Vol. (µL) !! Conc. (µg/mL) !! Amt (µg)
+|-
+| J23101 || 0.050 || 0.039 || 1.20 || 47 || 75 || 4.70
+|-
+| J23102 || 0.030 || 0.025 || 1.20 || 58 || 45 || 3.48
+|-
+| J23103 || 0.044 || 0.032 || 1.38 || 56 || 66 || 4.93
+|-
+| J23104 || 0.033 || 0.028 || 1.18 || 54 || 49 || 3.56
+|-
+| J23105 || 0.037 || 0.029 || 1.28 || 32 || 56 || 1.78
+|-
+| J23107 || 0.054 || 0.040 || 1.35 || 47 || 81 || 3.81
+|}
 [[User:Trentay|Trentay]] 13:44, 14 July 2009 (UTC)
+* Gel Electrophoresis Results
+[[Image:2009-07-14 digest06 100-105,107.jpg|350px|thumb|none|from left to right: ladder, 100-105,107]]
 ''Kevin and Adam''
 *Miniprep results:
+{| cellpadding="5" border="1" padding="5"
+|+
+|-
+! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !!  Vol. (µL) !! Conc. (µg/mL) !! Amt (µg)
+|-
+|E0240||.033||.019||1.737||124||66||8.184
+|-
+|pSB4C5||.054||.036||1.5||123.5||108||13.34
+|}
 [[User:Bussingkm|Bussingkm]] 15:21, 14 July 2009 (UTC)
 ''Craig and Clay''
-* The ampicillin plates left overnight again displayed no growth.
+* The ampicillin plates left overnight again displayed no growth.  The electrotransformed parts will be re-plated onto ampicillin-selective plates as well as non-selective plates.
-* The non-selective plates showed lawn growth.
 ----
@@ Line 67: / Line 103: @@
 ''Kevin and Adam''
+*perform complete digestion and ligation
 *construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106)
 [[User:Bussingkm|Bussingkm]] 15:20, 14 July 2009 (UTC)
 ''Craig and Clay''
-* The cells grown on the non-selective media will be streaked onto plates containing kanamycin and chloramphenicol to rule out possible mislabeling of resistance.
+* The electrotransformed parts will be re-plated on ampicillin plates as well as on non-selective plates.  Growth on non-selective plates will prove the existence of viable cells.
 ----
 ====Wetlab====
@@ Line 90: / Line 127: @@
 ''Craig and Clay''
-* The cells grown on the non-selective plates were streaked onto kanamycin and chloramphenicol plates.
+* The electrotransformed parts were again plated on ampicillin-selective plates.  They were also plated on non-selective media.  One hundred microliters was used for each plate rather than the standard 35 microliters.
-* Plates containing tetracycline need to be prepared to rule out this resistance.

Virginia Commonwealth/14 July 2009

From 2009.igem.org

Latest revision as of 20:45, 17 October 2009

Contents

Tuesday 14 July 2009

Results

Tasks

Wetlab