Team:Calgary/16 July 2009

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University of Calgary

THIS MONTH

July
M	T	W	T	F	S	S
		[http://2009.igem.org/Team:Calgary/1_July_2009 1]	[http://2009.igem.org/Team:Calgary/2_July_2009 2]	[http://2009.igem.org/Team:Calgary/3_July_2009 3]	[http://2009.igem.org/Team:Calgary/4_July_2009 4]	[http://2009.igem.org/Team:Calgary/5_July_2009 5]
[http://2009.igem.org/Team:Calgary/6_July_2009 6]	[http://2009.igem.org/Team:Calgary/7_July_2009 7]	[http://2009.igem.org/Team:Calgary/8_July_2009 8]	[http://2009.igem.org/Team:Calgary/9_July_2009 9]	[http://2009.igem.org/Team:Calgary/10_July_2009 10]	[http://2009.igem.org/Team:Calgary/11_July_2009 11]	[http://2009.igem.org/Team:Calgary/12_July_2009 12]
[http://2009.igem.org/Team:Calgary/13_July_2009 13]	[http://2009.igem.org/Team:Calgary/14_July_2009 14]	[http://2009.igem.org/Team:Calgary/15_July_2009 15]	[http://2009.igem.org/Team:Calgary/16_July_2009 16]	[http://2009.igem.org/Team:Calgary/17_July_2009 17]	[http://2009.igem.org/Team:Calgary/18_July_2009 18]	[http://2009.igem.org/Team:Calgary/19_July_2009 19]
[http://2009.igem.org/Team:Calgary/20_July_2009 20]	[http://2009.igem.org/Team:Calgary/21_July_2009 21]	[http://2009.igem.org/Team:Calgary/22_July_2009 22]	[http://2009.igem.org/Team:Calgary/23_July_2009 23]	[http://2009.igem.org/Team:Calgary/24_July_2009 24]	[http://2009.igem.org/Team:Calgary/25_July_2009 25]	[http://2009.igem.org/Team:Calgary/26_July_2009 26]
[http://2009.igem.org/Team:Calgary/27_July_2009 27]	[http://2009.igem.org/Team:Calgary/28_July_2009 28]	[http://2009.igem.org/Team:Calgary/29_July_2009 29]	[http://2009.igem.org/Team:Calgary/30_July_2009 30]	[http://2009.igem.org/Team:Calgary/31_July_2009 31]

NOTEBOOK PAGE INDEX

Carol
Chinmoyee
Emily
Fahd
Iman
Jamie
Jeremy
Katie
Kevin
Mandy
Patrick
Prima
Stefan
Vicki

CALENDAR

May
M	T	W	T	F	S	S
				[http://2009.igem.org/Team:Calgary/1_May_2009 1]	[http://2009.igem.org/Team:Calgary/2_May_2009 2]	[http://2009.igem.org/Team:Calgary/3_May_2009 3]
[http://2009.igem.org/Team:Calgary/4_May_2009 4]	[http://2009.igem.org/Team:Calgary/5_May_2009 5]	[http://2009.igem.org/Team:Calgary/6_May_2009 6]	[http://2009.igem.org/Team:Calgary/7_May_2009 7]	[http://2009.igem.org/Team:Calgary/8_May_2009 8]	[http://2009.igem.org/Team:Calgary/9_May_2009 9]	[http://2009.igem.org/Team:Calgary/10_May_2009 10]
[http://2009.igem.org/Team:Calgary/11_May_2009 11]	[http://2009.igem.org/Team:Calgary/12_May_2009 12]	[http://2009.igem.org/Team:Calgary/13_May_2009 13]	[http://2009.igem.org/Team:Calgary/14_May_2009 14]	[http://2009.igem.org/Team:Calgary/15_May_2009 15]	[http://2009.igem.org/Team:Calgary/16_May_2009 16]	[http://2009.igem.org/Team:Calgary/17_May_2009 17]
[http://2009.igem.org/Team:Calgary/18_May_2009 18]	[http://2009.igem.org/Team:Calgary/19_May_2009 19]	[http://2009.igem.org/Team:Calgary/20_May_2009 20]	[http://2009.igem.org/Team:Calgary/21_May_2009 21]	[http://2009.igem.org/Team:Calgary/22_May_2009 22]	[http://2009.igem.org/Team:Calgary/23_May_2009 23]	[http://2009.igem.org/Team:Calgary/24_May_2009 24]
[http://2009.igem.org/Team:Calgary/25_May_2009 25]	[http://2009.igem.org/Team:Calgary/26_May_2009 26]	[http://2009.igem.org/Team:Calgary/27_May_2009 27]	[http://2009.igem.org/Team:Calgary/28_May_2009 28]	[http://2009.igem.org/Team:Calgary/29_May_2009 29]	[http://2009.igem.org/Team:Calgary/30_May_2009 30]	[http://2009.igem.org/Team:Calgary/31_May_2009 31]

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

July
M	T	W	T	F	S	S
		[http://2009.igem.org/Team:Calgary/1_July_2009 1]	[http://2009.igem.org/Team:Calgary/2_July_2009 2]	[http://2009.igem.org/Team:Calgary/3_July_2009 3]	[http://2009.igem.org/Team:Calgary/4_July_2009 4]	[http://2009.igem.org/Team:Calgary/5_July_2009 5]
[http://2009.igem.org/Team:Calgary/6_July_2009 6]	[http://2009.igem.org/Team:Calgary/7_July_2009 7]	[http://2009.igem.org/Team:Calgary/8_July_2009 8]	[http://2009.igem.org/Team:Calgary/9_July_2009 9]	[http://2009.igem.org/Team:Calgary/10_July_2009 10]	[http://2009.igem.org/Team:Calgary/11_July_2009 11]	[http://2009.igem.org/Team:Calgary/12_July_2009 12]
[http://2009.igem.org/Team:Calgary/13_July_2009 13]	[http://2009.igem.org/Team:Calgary/14_July_2009 14]	[http://2009.igem.org/Team:Calgary/15_July_2009 15]	[http://2009.igem.org/Team:Calgary/16_July_2009 16]	[http://2009.igem.org/Team:Calgary/17_July_2009 17]	[http://2009.igem.org/Team:Calgary/18_July_2009 18]	[http://2009.igem.org/Team:Calgary/19_July_2009 19]
[http://2009.igem.org/Team:Calgary/20_July_2009 20]	[http://2009.igem.org/Team:Calgary/21_July_2009 21]	[http://2009.igem.org/Team:Calgary/22_July_2009 22]	[http://2009.igem.org/Team:Calgary/23_July_2009 23]	[http://2009.igem.org/Team:Calgary/24_July_2009 24]	[http://2009.igem.org/Team:Calgary/25_July_2009 25]	[http://2009.igem.org/Team:Calgary/26_July_2009 26]
[http://2009.igem.org/Team:Calgary/27_July_2009 27]	[http://2009.igem.org/Team:Calgary/28_July_2009 28]	[http://2009.igem.org/Team:Calgary/29_July_2009 29]	[http://2009.igem.org/Team:Calgary/30_July_2009 30]	[http://2009.igem.org/Team:Calgary/31_July_2009 31]

August
M	T	W	T	F	S	S
					[http://2009.igem.org/Team:Calgary/1_August_2009 1]	[http://2009.igem.org/Team:Calgary/2_August_2009 2]
[http://2009.igem.org/Team:Calgary/3_August_2009 3]	[http://2009.igem.org/Team:Calgary/4_August_2009 4]	[http://2009.igem.org/Team:Calgary/5_August_2009 5]	[http://2009.igem.org/Team:Calgary/6_August_2009 6]	[http://2009.igem.org/Team:Calgary/7_August_2009 7]	[http://2009.igem.org/Team:Calgary/8_August_2009 8]	[http://2009.igem.org/Team:Calgary/9_August_2009 9]
[http://2009.igem.org/Team:Calgary/10_August_2009 10]	[http://2009.igem.org/Team:Calgary/11_August_2009 11]	[http://2009.igem.org/Team:Calgary/12_August_2009 12]	[http://2009.igem.org/Team:Calgary/13_August_2009 13]	[http://2009.igem.org/Team:Calgary/14_August_2009 14]	[http://2009.igem.org/Team:Calgary/15_August_2009 15]	[http://2009.igem.org/Team:Calgary/16_August_2009 16]
[http://2009.igem.org/Team:Calgary/17_August_2009 17]	[http://2009.igem.org/Team:Calgary/18_August_2009 18]	[http://2009.igem.org/Team:Calgary/19_August_2009 19]	[http://2009.igem.org/Team:Calgary/20_August_2009 20]	[http://2009.igem.org/Team:Calgary/21_August_2009 21]	[http://2009.igem.org/Team:Calgary/22_August_2009 22]	[http://2009.igem.org/Team:Calgary/23_August_2009 23]
[http://2009.igem.org/Team:Calgary/24_August_2009 24]	[http://2009.igem.org/Team:Calgary/25_August_2009 25]	[http://2009.igem.org/Team:Calgary/26_August_2009 26]	[http://2009.igem.org/Team:Calgary/27_August_2009 27]	[http://2009.igem.org/Team:Calgary/28_August_2009 28]	[http://2009.igem.org/Team:Calgary/29_August_2009 29]	[http://2009.igem.org/Team:Calgary/30_August_2009 30]
[http://2009.igem.org/Team:Calgary/31_August_2009 31]

September
M	T	W	T	F	S	S
	[http://2009.igem.org/Team:Calgary/1_September_2009 1]	[http://2009.igem.org/Team:Calgary/2_September_2009 2]	[http://2009.igem.org/Team:Calgary/3_September_2009 3]	[http://2009.igem.org/Team:Calgary/4_September_2009 4]	[http://2009.igem.org/Team:Calgary/5_September_2009 5]	[http://2009.igem.org/Team:Calgary/6_September_2009 6]
[http://2009.igem.org/Team:Calgary/7_September_2009 7]	[http://2009.igem.org/Team:Calgary/8_September_2009 8]	[http://2009.igem.org/Team:Calgary/9_September_2009 9]	[http://2009.igem.org/Team:Calgary/10_September_2009 10]	[http://2009.igem.org/Team:Calgary/11_September_2009 11]	[http://2009.igem.org/Team:Calgary/12_September_2009 12]	[http://2009.igem.org/Team:Calgary/13_September_2009 13]
[http://2009.igem.org/Team:Calgary/14_September_2009 14]	[http://2009.igem.org/Team:Calgary/15_September_2009 15]	[http://2009.igem.org/Team:Calgary/16_September_2009 16]	[http://2009.igem.org/Team:Calgary/17_September_2009 17]	[http://2009.igem.org/Team:Calgary/18_September_2009 18]	[http://2009.igem.org/Team:Calgary/19_September_2009 19]	[http://2009.igem.org/Team:Calgary/20_September_2009 20]
[http://2009.igem.org/Team:Calgary/21_September_2009 21]	[http://2009.igem.org/Team:Calgary/22_September_2009 22]	[http://2009.igem.org/Team:Calgary/23_September_2009 23]	[http://2009.igem.org/Team:Calgary/24_September_2009 24]	[http://2009.igem.org/Team:Calgary/25_September_2009 25]	[http://2009.igem.org/Team:Calgary/26_September_2009 26]	[http://2009.igem.org/Team:Calgary/27_September_2009 27]
[http://2009.igem.org/Team:Calgary/28_September_2009 28]	[http://2009.igem.org/Team:Calgary/29_September_2009 29]	[http://2009.igem.org/Team:Calgary/30_September_2009 30]

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

JULY 16, 2009

CAROL

Diluting Primers and Primer Extension Polymerase Chain Reaction (PCR)

To create the synthetic sigma 70 promoter library, we have synthesized 32 different primers (16 different forward primers and 16 different reverse primers) to hopefully form 256 different promoters.
Need to dilute primers to 80μM. The following is the concentration of the original primers that were synthesized once water was added to the primers.

Primer	260/280	260/230	Concentration [ng/μL]	Reading/2.5 [ng/μL]	Stock Concentration [μM]	Volume stock into working [μL]	Volume water into working [μL]
sig70-F1	1.98	1.73	20816	832.64	60.07	6.7	43.3
sig70-F2	1.55	1.80	44284	17713.6	1277.91	3.1	46.9
sig70-F3	1.62	1.86	41323	16529.2	1192.46	3.4	46.6
sig70-F4	1.70	1.99	32365	12946	933.96	4.3	45.7
sig70-F5	1.72	2.07	33688	13475.2	972.14	4.1	45.9
sig70-F6	2.07	1.95	45412	18164.8	1310.46	3.1	46.9
sig70-F7	1.64	1.95	39046	15618.4	1126.76	3.6	46.4
sig70-F8	1.43	1.63	45745	18298	1320.07	3.0	47.0
sig70-F9	1.75	2.16	23628	9451.2	681.84	5.9	44.1
sig70-F10	1.72	2.01	32682	13072.8	943.11	4.2	45.8
sig70-F11	1.71	2.07	31494	12597.6	908.83	4.4	45.6
sig70-F12	1.59	1.82	41202	16480.8	1188.97	3.4	46.6
sig70-F13	1.73	2.03	29224	11689.6	843.3	4.7	45.3
sig70-F14	1.41	1.58	45645	18258	1317.2	3.0	47.0
sig70-F15	1.59	1.85	40681	16272.4	1173.9	3.4	46.6
sig70-F16	1.57	1.82	42241	16896.4	1219.0	3.3	46.7
sig70-R1	2.06	2.17	7516	3006.4	216.9	18.4	31.6
sig70-R2	2.04	2.25	15255	6102.0	440.2	9.1	40.9
sig70-R3	1.76	1.85	42503	17001.2	1226.5	3.3	46.7
sig70-R4	2.01	2.21	24012	9604.8	692.9	5.8	44.2
sig70-R5	2.10	2.27	18530	7412	534.7	7.5	42.5
sig70-R6	1.98	2.19	28452	11380.8	821.0	4.9	45.1
sig70-R7	1.94	2.17	32976	13190.4	951.6	4.2	45.8
sig70-R8	2.00	2.20	25379	10151.6	732.4	5.5	44.5
sig70-R9	2.00	2.24	24141	9656.4	696.6	5.9	44.1
sig70-R10	1.94	2.17	30237	12094.8	872.6	4.6	45.4
sig70-R11	1.92	2.16	31497	12598.8	908.9	4.4	45.6
sig70-R12	1.92	2.17	30045	12018	867.0	4.6	45.4
sig70-R13	2.01	2.18	18726	7490.4	540.4	7.4	42.6
sig70-R14	1.88	2.03	36111	14444.4	1042.1	3.8	46.2
sig70-R15	1.98	2.17	10679	4271.6	308.2	13	37
sig70-R16	1.86	2.01	35969	14387.6	1038	3.9	46.1

The sequences for each primer is listed in the following table:

Primer	‘’’Sequence’’’
sig70-F1	GCGCGCTCGAGAATAATTCTTAAAATTTATGCTTCCGGCTCG
sig70-F2	GCGCGCTCGAGAATAATTCTTAATATTTATGCTTCCGGCTCG
sig70-F3	GCGCGCTCGAGAATAATTCTTAAGATTTATGCTTCCGGCTCG
sig70-F4	GCGCGCTCGAGAATAATTCTTAACATTTATGCTTCCGGCTCG
sig70-F5	GCGCGCTCGAGAATAATTCTTTAAATTTATGCTTCCGGCTCG
sig70-F6	GCGCGCTCGAGAATAATTCTTTATATTTATGCTTCCGGCTCG
sig70-F7	GCGCGCTCGAGAATAATTCTTTAGATTTATGCTTCCGGCTCG
sig70-F8	GCGCGCTCGAGAATAATTCTTTACATTTATGCTTCCGGCTCG
sig70-F9	GCGCGCTCGAGAATAATTCTTGAAATTTATGCTTCCGGCTCG
sig70-F10	GCGCGCTCGAGAATAATTCTTGATATTTATGCTTCCGGCTCG
sig70-F11	GCGCGCTCGAGAATAATTCTTGAGATTTATGCTTCCGGCTCG
sig70-F12	GCGCGCTCGAGAATAATTCTTGACATTTATGCTTCCGGCTCG
sig70-F13	GCGCGCTCGAGAATAATTCTTCAAATTTATGCTTCCGGCTCG
sig70-F14	GCGCGCTCGAGAATAATTCTTCATATTTATGCTTCCGGCTCG
sig70-F15	GCGCGCTCGAGAATAATTCTTCAGATTTATGCTTCCGGCTCG
sig70-F16	GCGCGCTCGAGAATAATTCTTCACATTTATGCTTCCGGCTCG
sig70-R1	GCGGGATCCAATTGCACGTAAAATACGAGCCGGAAGCATAAA
sig70-R2	GCGGGATCCAATTGCACGTAATATACGAGCCGGAAGCATAAA
sig70-R3	GCGGGATCCAATTGCACGTAACATACGAGCCGGAAGCATAAA
sig70-R4	GCGGGATCCAATTGCACGTAAGATACGAGCCGGAAGCATAAA
sig70-R5	GCGGGATCCAATTGCACGTATAATACGAGCCGGAAGCATAAA
sig70-R6	GCGGGATCCAATTGCACGTATTATACGAGCCGGAAGCATAAA
sig70-R7	GCGGGATCCAATTGCACGTATGATACGAGCCGGAAGCATAAA
sig70-R8	GCGGGATCCAATTGCACGTATCATACGAGCCGGAAGCATAAA
sig70-R9	GCGGGATCCAATTGCACGTAGAATACGAGCCGGAAGCATAAA
sig70-R10	GCGGGATCCAATTGCACGTAGTATACGAGCCGGAAGCATAAA
sig70-R11	GCGGGATCCAATTGCACGTAGGATACGAGCCGGAAGCATAAA
sig70-R12	GCGGGATCCAATTGCACGTAGCATACGAGCCGGAAGCATAAA
sig70-R13	GCGGGATCCAATTGCACGTACAATACGAGCCGGAAGCATAAA
sig70-R14	GCGGGATCCAATTGCACGTACTATACGAGCCGGAAGCATAAA
sig70-R15	GCGGGATCCAATTGCACGTACGATACGAGCCGGAAGCATAAA
sig70-R16	GCGGGATCCAATTGCACGTACCATACGAGCCGGAAGCATAAA

Stock dATP, dGTP, dCTP, dTTP from Invitrogen was diluted from 100mM to 0.5mM.
See how the primer extension PCR is done in the protocol page.
Both the varying promoters and the vector, pCS26, was digested using enzymes, XhoI and BamHI overnight. The vector was incubated at 37^oC and the promoter was digested for 2 hours at 16^oC and then overnight at room temperature (~21^oC).

CHINMOYEE