IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/01

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7/1/14 Gel Electrophoresis

  • Gel Electrophoresis of Plasmids
  • Made TAE buffer with 1 L of distilled water and 20mL of 50X stock of TAE. Also prepared the gel with agarose and water mixture and let solidify.
  • Made plasmids ready by using 3μL of DNA and 3μL of loading dye. 5 tubes of this were made as well as 1 tube of ladder 3μL.
  • Insert ladder as well as DNA plasmids in wells and let sit for around 45-50 minutes.
  • This was to determine concentration of plasmids
  • Next, the sizes of the plasmids were tested to see how accurate they are compared to the literature value
  • Used the plasmids and cut them with restriction digests
  • 2μL of DNA, 1μL of Buffer, 5μL of water, 2μL of, 1μL of E and 1μ of P restriction digests.
  • Heated in a 37°C water bath for around 40 minutes
  • Plasmids that have been cut were taken out and loading dye was input--2μL
  • This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V
  • Pictures were taken of both gels and data recorded.



Reagent Volume Expected:
1. TesA = 2721
2. mCherry = size
3. TetR = 2124
4. Lac = 2125
5. RBS= 2080

15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 2.0 μL
10X buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 9.5
  15 μL --> 37°C/ ~40 min.


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{"cc_re__cc_re__cc_by_sa_2_5": ["a href=\"/wiki/OpenWetWare:How_to_join\">\n\t\t\t<img style=\"margin:2px;\" \n\t\t\t\tsrc=\"http://openwetware.org/images/0/03/88x31_JoinOWW.png\" border=0 /></a>\n                </div>\n        </div>\n\t\t\t\t\t\t\t</div>\n\n\t\t\t</div><!-- end of the left column -->\n\t\t\n\t\t\n\t\t\t<div class=\"visualClear\"></div>\n\t\t\t<div id=\"footer\">\n\t\t\t\t<div id=\"f-poweredbyico\"><a href=\"http://www.mediawiki.org/\"><img src=\"/skins/common/images/poweredby_mediawiki_88x31.png\" alt=\"Powered by MediaWiki\" /></a></div>\n\t\t\t\t<div id=\"f-copyrightico\"><a href=\"http://creativecommons.org/licenses/by-sa/2.5/\"><img src=\"/somerights20.png\" alt='GNU FDL or Creative Commons BY-SA' /></a></div>\n\t\t\t<ul id=\"f-list\">\n\t\t\t\t<li id=\"copyright\">Content is available under <a href=\"/wiki/Copyright\" title=\"Copyright\">GNU FDL or Creative Commons BY-SA</a>.</li>\n\t\t\t\t<li id=\"privacy\"><a href=\"/wiki/OpenWetWare:Privacy_policy\" title=\"OpenWetWare:Privacy policy\">Privacy policy</a></li>\n\t\t\t\t<li id=\"about\"><a href=\"/wiki/OpenWetWare:About\" title=\"OpenWetWare:About\">About OpenWetWare</a></li>\n\t\t\t\t<li id=\"disclaimer\"><a href"], "copyright__copyright__copyright": ["een cut were taken out and loading dye was input--2\u03bcL\n  \u2022 This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V\n  \u2022 Pictures were taken of both gels and data recorded.\n\n\n\nReagent Volume Expected:\n1. TesA = 2721\n2. mCherry = size\n3. TetR = 2124\n4. Lac = 2125\n5. RBS= 2080\n\n15 \u03bcL/lane; 1% agarose; Ladder\nDNA(plasmid) 2.0 \u03bcL\n10X buffer 1.5\nEcoRI 1.0\nPstI 1.0\ndH2O 9.5\n\u00a0 15 \u03bcL --> 37\u00b0C/ ~40 min.\n\n\nPersonal tools", "een cut were taken out and loading dye was input--2\u03bcL\n  \u2022 This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V\n  \u2022 Pictures were taken of both gels and data recorded.\n\n\n\nReagent Volume Expected:\n1. TesA = 2721\n2. mCherry = size\n3. TetR = 2124\n4. Lac = 2125\n5. RBS= 2080\n\n15 \u03bcL/lane; 1% agarose; Ladder\nDNA(plasmid) 2.0 \u03bcL\n10X buffer 1.5\nEcoRI 1.0\nPstI 1.0\ndH2O 9.5\n\u00a0 15 \u03bcL --> 37\u00b0C/ ~40 min.\n\n\nPersonal tools", "een cut were taken out and loading dye was input--2\u03bcL\n  \u2022 This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V\n  \u2022 Pictures were taken of both gels and data recorded.\n\n\n\nReagent Volume Expected:\n1. TesA = 2721\n2. mCherry = size\n3. TetR = 2124\n4. Lac = 2125\n5. RBS= 2080\n\n15 \u03bcL/lane; 1% agarose; Ladder\nDNA(plasmid) 2.0 \u03bcL\n10X buffer 1.5\nEcoRI 1.0\nPstI 1.0\ndH2O 9.5\n\u00a0 15 \u03bcL --> 37\u00b0C/ ~40 min.\n\n\nPersonal tools", "een cut were taken out and loading dye was input--2\u03bcL\n  \u2022 This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V\n  \u2022 Pictures were taken of both gels and data recorded.\n\n\n\nReagent Volume Expected:\n1. TesA = 2721\n2. mCherry = size\n3. TetR = 2124\n4. Lac = 2125\n5. RBS= 2080\n\n15 \u03bcL/lane; 1% agarose; Ladder\nDNA(plasmid) 2.0 \u03bcL\n10X buffer 1.5\nEcoRI 1.0\nPstI 1.0\ndH2O 9.5\n\u00a0 15 \u03bcL --> 37\u00b0C/ ~40 min.\n\n\nPersonal tools", "een cut were taken out and loading dye was input--2\u03bcL\n  \u2022 This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V\n  \u2022 Pictures were taken of both gels and data recorded.\n\n\n\nReagent Volume Expected:\n1. TesA = 2721\n2. mCherry = size\n3. TetR = 2124\n4. Lac = 2125\n5. RBS= 2080\n\n15 \u03bcL/lane; 1% agarose; Ladder\nDNA(plasmid) 2.0 \u03bcL\n10X buffer 1.5\nEcoRI 1.0\nPstI 1.0\ndH2O 9.5\n\u00a0 15 \u03bcL --> 37\u00b0C/ ~40 min.\n\n\nPersonal tools", "een cut were taken out and loading dye was input--2\u03bcL\n  \u2022 This along with the ladder was filled into the wells and the machine was run for around 35 minutes. The voltage was increased from 80 V to 100 V\n  \u2022 Pictures were taken of both gels and data recorded.\n\n\n\nReagent Volume Expected:\n1. TesA = 2721\n2. mCherry = size\n3. TetR = 2124\n4. Lac = 2125\n5. RBS= 2080\n\n15 \u03bcL/lane; 1% agarose; Ladder\nDNA(plasmid) 2.0 \u03bcL\n10X buffer 1.5\nEcoRI 1.0\nPstI 1.0\ndH2O 9.5\n\u00a0 15 \u03bcL --> 37\u00b0C/ ~40 min.\n\n\nPersonal tools"]}