From 2010.igem.org
August 23
Today was a productive day.
We prepared 50 ml of Poliacrilamid gel 8% with 0.5 x TBE.
- 13,3 ml ABA (Arilamid - Bisacrilamid)
- 2,5 ml of TBE 10 x
- 34,2 ml of H2O
- 50 ul of APS 25%
- 50 ul of TEMED
Then we seal the borders with 2% of agarose, fill with 0,5 TBE x 50 ml and
Reveal with 5 00 ml H2O and 25 ul o Bromure x 20 minutes.
10-15 ul of sample
150 V and 17 mamps
12 %
- 19,95 ml ABA 10 ml
- 3,75 ml TBE 10x 1,87 ml
- 75 ul APS 37,5 ul
- 75 ul TE MED 37,5 ul
- 26, 3 ml H2O 13, 15 ul
After all of these, we finally, started de ligation of RBS, promotor and plasmid.
The ligation mix that we use was:
- 11 ul H2O
- 2 ul digested RBS
- 2 ul digested Promotor
- 2 ul digested plasmid
- 2 ul T4 10x ligase buffer
- 1 ul T4 ligase
So to get a final volume of 20 ul, then we incubated this for 2 hours at room temperature
and later at 80ºC for 20 minutes to desactivate the enzime.
And good news for everyone, the ligation was done right!
so now we have our RBS and promotor in a plasmid =)
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/12_May_2010&action=edit 12]
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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/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]
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[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]
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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]
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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/Team:Panama/26_June_2010 26]
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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]
| [http://2010.igem.org/wiki/index.php?title=Team:Panama/25_July_2010&action=edit 25]
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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]
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| [http://2010.igem.org/Team:Panama/19_August_2010 19]
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| [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]
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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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