Team:Panama/28 July 2010

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Agarose gel:
Agarose gel:
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Once the gel was set, in each well we added 20 ul of sample, plus 2ul of loading buffer. Total volume = 22ul.
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Once the gel was set, in each well we load 20 ul of sample, plus 2ul of loading buffer. Total volume = 22ul.
The molecular weight marker that we used was the 25 bp DNA step ladder, G451A.
The molecular weight marker that we used was the 25 bp DNA step ladder, G451A.
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Results: We could visualize the DNA bands on the agarose gel.  
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Results:  
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We could visualize the DNA bands on the agarose gel.  

Revision as of 20:02, 24 October 2010

iGEM Panama

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July 28

Electrophoresis of the digested parts:

4% 3:1 Agarose (Amresco, Cat no: 9012-36-6)

100ml TBE 1X.

4g of agarose

The agarose is disolved in the TBE using heat. At first, the mixture is opaque but as the agarose disolves, the mixture becomes clear. At this point we add 5ul of Ethydium bromide. This can then be poured into the casting chamber.

Agarose gel:

Once the gel was set, in each well we load 20 ul of sample, plus 2ul of loading buffer. Total volume = 22ul.

The molecular weight marker that we used was the 25 bp DNA step ladder, G451A.

Application sample order on the agarose gel: M (for the marker), 2 (plasmid), 3 (promoter) & 4 (RBS)


Results: We could visualize the DNA bands on the agarose gel.


Picture






Transformation of E. coli using the part terminator. For this transformation we followed the BioBrick protocol. We took 2 ul of the resuspended DNA stock that was stored at -20 and did a transformation with heat shock.

We tried to run another gel with a different concentration of agarose to see if the reason that we can't see the insert bands was because of the gel.

Agarose Gel 2% (3:1) Amresco -100 ml TBE 1x -2 g Agarose -Dissolve


  • Picture





The restriction digestions were not successful, here are the possible reasons:

1) Exists the possibility that the enzymes use for the restriction do not work adequately (possibly weren't kept at the correct temperature during shipment). "The digestion did not occur"

2) The molecular weight marker that we used (Amresco G451A 25bp) recommends that to be able to visualize bands between 25-50 bp, you should add 1.8 ug, or 1800 ng, into the well in order to visualize the bands. In the case of the marker they recommend 5 ul.

Miniprep As a result of our plasmid DNA miniprep we obtained 30ng/ul. For the restriction reaction we used 500ng of plasmid DNA, therefore 20 ul of the miniprep.

Then from the digestion we obtained a volume of 50 ul. From here we took 20 ul to load into a gel. This means that the real quantity of DNA that we loaded into the gel was 240 ng.

For this reason it is possible that there wasn't enough DNA (1800ng vs. 240 ng) to be able to see the bands.

"The digestion occurred but we weren't able to see the small DNA bands"

May
MTWTFSS
          [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
MTWTFSS
  [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
MTWTFSS
      [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
MTWTFSS
            [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
MTWTFSS
    [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
MTWTFSS
        [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]