From 2010.igem.org
(Difference between revisions)
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==='''October 13'''===
==='''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:
+
+ ''' Quick Screening Protocol:'''
100ul of overnight culture
100ul of overnight culture
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10ul Loading Buffer
10ul Loading Buffer
- We loaded 20ul into each well
+ We mix all the reaction component with a vortex, and then we loaded 20ul into each well of a 1% agarose gel.
[[Image:Cuadro13octubre2.jpg|500px|thumb|left|alt text]]
[[Image:Cuadro13octubre2.jpg|500px|thumb|left|alt text]]
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- '''Preparation of the Primers for Mutagenesis'''
+ '''Stock solution preparation of the Primers for Mutagenesis'''
RhT-g 159 a
RhT-g 159 a
Revision as of 03:32, 28 October 2010
iGEM Panama
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
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_May_2010&action=edit 1 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/2_May_2010&action=edit 2 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/3_May_2010&action=edit 3 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/4_May_2010&action=edit 4 ]
[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 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/9_May_2010&action=edit 9 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/10_May_2010&action=edit 10 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/11_May_2010&action=edit 11 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/12_May_2010&action=edit 12 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/13_May_2010&action=edit 13 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/14_May_2010&action=edit 14 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/15_May_2010&action=edit 15 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/16_May_2010&action=edit 16 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/17_May_2010&action=edit 17 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_May_2010&action=edit 18 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_May_2010&action=edit 19 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/20_May_2010&action=edit 20 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_May_2010&action=edit 21 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_May_2010&action=edit 22 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/23_May_2010&action=edit 23 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/24_May_2010&action=edit 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_May_2010&action=edit 25 ]
[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 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/31_May_2010&action=edit 31 ]
June
M T W T F S S
[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 ]
[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 ]
[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 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/18_June_2010&action=edit 18 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/19_June_2010&action=edit 19 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/20_June_2010&action=edit 20 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/21_June_2010&action=edit 21 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/22_June_2010&action=edit 22 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/23_June_2010&action=edit 23 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/24_June_2010&action=edit 24 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_June_2010&action=edit 25 ]
[http://2010.igem.org/Team:Panama/26_June_2010 26 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/27_June_2010&action=edit 27 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/28_June_2010&action=edit 28 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/29_June_2010&action=edit 29 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/30_June_2010&action=edit 30 ]
July
M T W T F S S
[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 ]
[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 ]
[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 ]
[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 ]
[http://2010.igem.org/wiki/index.php?title=Team:Panama/25_July_2010&action=edit 25 ]
[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 ]
August
M T W T F S S
[http://2010.igem.org/wiki/index.php?title=Team:Panama/1_August_2010&action=edit 1 ]
[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 ]
[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 ]
[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 ]
[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 ]
[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 ]
September
M T W T F S S
[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 ]
[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 ]
[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 ]
[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 ]
[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 ]
October
M T W T F S S
[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 ]
[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 ]
[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 ]
[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 ]
[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 ]