October 13
In order to save time and expensive reagents we did a Quick check screening protocol, this experiment will help us find a colony with the transformed bacteria.
Quick Screening Protocol:
100ul of overnight culture
50ul Phenol:Chloroform:Isoamyl alcohol (25:24:1)
10ul Loading Buffer
We mix all the reaction component with a vortex, and then we loaded 20ul into each well of a 1% agarose gel.
Stock solution preparation of the Primers for Mutagenesis
RhT-g 159 a
1) 5' CGA ATC ACG TGC TAC AGC GCG CCT ACG 3'
Resuspend the lyophilized primers for stock soluction (100 uM)
27.9 nM x 10 = 279 ul of water
Working solution (10 uM)
10 ul of stock + 90 ul water
2) RhT-C62T
5' ACC TGC CGC ACT GTA GTT TCT CGC GGG 3'
33.4 nM x 10 = 334 ul of water
Working solution (10 uM)
10 ul of stock + 90 ul of water
3) RhT-g45a
5' CTC GAC CGA ACA CCT ACA GGG CGA CTT CTA C 3'
32.1 nM x 10 = 321 ul of water
Working solution (10 uM)
10 ul of stock + 90 ul of water
We did the miniprep of the cultures, wells 4 and 5
3, 4, 5 and 7 seem to be bigger
We did a PCR following the guidelines on pg. 53 to verify if the cultures of well 4 and 5 actually have the gene Rh1ab so that we can continue with the mutagenesis.
May
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_May_2010&action=edit 1]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_May_2010&action=edit 3]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_May_2010&action=edit 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_May_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_May_2010&action=edit 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_May_2010&action=edit 8]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/10_May_2010&action=edit 10]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_May_2010&action=edit 17]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/24_May_2010&action=edit 24]
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| [http://2010.igem.org/Team:Panama/26_May_2010 26]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/27_May_2010&action=edit 27]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_May_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_May_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_May_2010&action=edit 30]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_May_2010&action=edit 31]
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June
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_June_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_June_2010&action=edit 2]
| [http://2010.igem.org/Team:Panama/3_June_2010 3]
| [http://2010.igem.org/Team:Panama/4_June_2010 4]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_June_2010&action=edit 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_June_2010&action=edit 6]
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[http://2010.igem.org/Team:Panama/7_June_2010 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_June_2010&action=edit 8]
| [http://2010.igem.org/Team:Panama/9_June_2010 9]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_June_2010&action=edit 10]
| [http://2010.igem.org/Team:Panama/11_June_2010 11]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/12_June_2010&action=edit 12]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/13_June_2010&action=edit 13]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_June_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_June_2010&action=edit 15]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/16_June_2010&action=edit 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_June_2010&action=edit 17]
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/19_June_2010&action=edit 19]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_June_2010&action=edit 21]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_June_2010&action=edit 28]
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July
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_July_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_July_2010&action=edit 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_July_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_July_2010&action=edit 4]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/5_July_2010&action=edit 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_July_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_July_2010&action=edit 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_July_2010&action=edit 8]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/9_July_2010&action=edit 9]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_July_2010&action=edit 10]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/11_July_2010&action=edit 11]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_July_2010&action=edit 12]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/13_July_2010&action=edit 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_July_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_July_2010&action=edit 15]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/16_July_2010&action=edit 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_July_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_July_2010&action=edit 18]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_July_2010&action=edit 19]
| [http://2010.igem.org/Team:Panama/20_July_2010 20]
| [http://2010.igem.org/Team:Panama/21_July_2010 21]
| [http://2010.igem.org/Team:Panama/22_July_2010 22]
| [http://2010.igem.org/Team:Panama/23_July_2010 23]
| [http://2010.igem.org/Team:Panama/24_July_2010 24]
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[http://2010.igem.org/Team:Panama/26_July_2010 26]
| [http://2010.igem.org/Team:Panama/27_July_2010 27]
| [http://2010.igem.org/Team:Panama/28_July_2010 28]
| [http://2010.igem.org/Team:Panama/29_July_2010 29]
| [http://2010.igem.org/Team:Panama/30_July_2010 30]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_July_2010&action=edit 31]
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August
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_August_2010&action=edit 1]
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[http://2010.igem.org/Team:Panama/2_August_2010 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_August_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_August_2010&action=edit 4]
| [http://2010.igem.org/Team:Panama/5_August_2010 5]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/6_August_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_August_2010&action=edit 7]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/8_August_2010&action=edit 8]
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[http://2010.igem.org/Team:Panama/9_August_2010 9]
| [http://2010.igem.org/Team:Panama/10_August_2010 10]
| [http://2010.igem.org/Team:Panama/11_August_2010 11]
| [http://2010.igem.org/Team:Panama/12_August_2010 12]
| [http://2010.igem.org/Team:Panama/13_August_2010 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_August_2010&action=edit 14]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/15_August_2010&action=edit 15]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_August_2010&action=edit 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_August_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_August_2010&action=edit 18]
| [http://2010.igem.org/Team:Panama/19_August_2010 19]
| [http://2010.igem.org/Team:Panama/20_August_2010 20]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/21_August_2010&action=edit 21]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/22_August_2010&action=edit 22]
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[http://2010.igem.org/Team:Panama/23_August_2010 23]
| [http://2010.igem.org/Team:Panama/24_August_2010 24]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_August_2010&action=edit 25]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/26_August_2010&action=edit 26]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/27_August_2010&action=edit 27]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_August_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_August_2010&action=edit 29]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_August_2010&action=edit 30]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_August_2010&action=edit 31]
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September
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_September_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_September_2010&action=edit 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_September_2010&action=edit 3]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/4_September_2010&action=edit 4]
| [http://2010.igem.org/Team:Panama/5_September_2010 5]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/6_September_2010&action=edit 6]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/7_September_2010&action=edit 7]
| [http://2010.igem.org/Team:Panama/8_September_2010 8]
| [http://2010.igem.org/Team:Panama/9_September_2010 9]
| [http://2010.igem.org/Team:Panama/10_September_2010 10]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/11_September_2010&action=edit 11]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/12_September_2010&action=edit 12]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/13_September_2010&action=edit 13]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/14_September_2010&action=edit 14]
| [http://2010.igem.org/Team:Panama/15_September_2010 15]
| [http://2010.igem.org/Team:Panama/16_September_2010 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_September_2010&action=edit 17]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/18_September_2010&action=edit 18]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/19_September_2010&action=edit 19]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/20_September_2010&action=edit 20]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/21_September_2010&action=edit 21]
| [http://2010.igem.org/Team:Panama/22_September_2010 22]
| [http://2010.igem.org/Team:Panama/23_September_2010 23]
| [http://2010.igem.org/Team:Panama/24_September_2010 24]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_September_2010&action=edit 25]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/26_September_2010&action=edit 26]
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[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_September_2010&action=edit 27]
| [http://2010.igem.org/Team:Panama/28_September_2010 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_September_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_September_2010&action=edit 30]
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October
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| [http://2010.igem.org/wiki/index.php?title=Team:Panama/1_October_2010&action=edit 1]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/2_October_2010&action=edit 2]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/3_October_2010&action=edit 3]
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[http://2010.igem.org/Team:Panama/4_October_2010 4]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/5_October_2010&action=edit 5]
| [http://2010.igem.org/Team:Panama/6_October_2010 6]
| [http://2010.igem.org/Team:Panama/7_October_2010 7]
| [http://2010.igem.org/Team:Panama/8_October_2010 8]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/9_October_2010&action=edit 9]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/10_October_2010&action=edit 10]
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[http://2010.igem.org/Team:Panama/11_October_2010 11]
| [http://2010.igem.org/Team:Panama/12_October_2010 12]
| [http://2010.igem.org/Team:Panama/13_October_2010 13]
| [http://2010.igem.org/Team:Panama/14_October_2010 14]
| [http://2010.igem.org/Team:Panama/15_October_2010 15]
| [http://2010.igem.org/Team:Panama/16_October_2010 16]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/17_October_2010&action=edit 17]
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[http://2010.igem.org/Team:Panama/18_October_2010 18]
| [http://2010.igem.org/Team:Panama/19_October_2010 19]
| [http://2010.igem.org/Team:Panama/20_October_2010 20]
| [http://2010.igem.org/Team:Panama/21_October_2010 21]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/22_October_2010&action=edit 22]
| [http://2010.igem.org/Team:Panama/23_October_2010 23]
| [http://2010.igem.org/Team:Panama/24_October_2010 24]
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[http://2010.igem.org/Team:Panama/25_October_2010 25]
| [http://2010.igem.org/Team:Panama/26_October_2010 26]
| [http://2010.igem.org/Team:Panama/27_October_2010 27]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/28_October_2010&action=edit 28]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/29_October_2010&action=edit 29]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/30_October_2010&action=edit 30]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/31_October_2010&action=edit 31]
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