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| - | <h1 class="title"> Protocols //Notebook </h1>
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| - | <div class="book_wrapper"> <a id="next_page_button"></a> <a id="prev_page_button"></a>
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| - | <div class="loading" id="loading">Loading pages...</div>
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| - | <div style="display:none;" id="mybook">
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| - | <div> <img alt="" src='https://static.igem.org/mediawiki/2014hs/8/88/FotoiGEM1.jpg' width='400' height='300' />
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| - | <h1>Protocol 1 </h1>
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| - | <p>Preparation of culture media (1L).<br />
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| - | 1.- 5 g of yeast<br />
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| - | 2.- 10 g of sodium chloride.<br />
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| - | 3.- 10 g of triptone.<br />
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| - | 4.- Add water till 1 uL volume.</p>
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| - | </div>
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| - | <div>
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| - | <h1>Protocol 2</h1>
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| - | <p>Competent cell´s preparation<br />
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| - | <br />
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| - | 1.- Centrifug your media culture tube (25 mL) at 4,000 rpm during
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| - | 15 min.<br />
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| - | 2.- Pour the supernatant and add 10 mL of cold MgCl2 solution.
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| - | Centrifuge them at 4,000 rpm at 4 °C during 15 min.<br />
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| - | 3.- Pour the supernatant and resuspend the pellet in 40 mL of cold
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| - | CaCl2 solution. Keep it in ice during 20 min and place them in
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| - | eppendorf tubes. Centrifuge them at 4,000 rpm at 4°C during 15
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| - | min.<br />
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| - | 4.- Pour the supernatant and resuspend the pellet in 40 mL of cold
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| - | 85mM CaCl2/15% glycerlol v/v solution. Centrifuge again at 2,100
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| - | rpm at 4°C during 15 min.<br />
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| - | 5.- Pour the supernatant and resuspend in 2 mL of cold 85bmM
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| - | CaCl2/15% glycerol v/v solution.<br />
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| - | 6.- Aliquote in tubes of 100 uL previously frozen and store at
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| - | -80°C.<br />
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| - | </p>
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| - | </div>
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| - | <div>
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| - | <h1>Protocol 3</h1>
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| - | <p>DNA extraction from Buffy coat:<br />
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| - | <br />
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| - | <font size="1"> 1.- Centrifuge 5 mL of culture medium at 1,500 rpm
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| - | for 10 min at 4°C.</font><br />
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| - | <font size="1">2.- Carefully transfer 600 uL of buffy coat to a
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| - | microtube.</font><br />
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| - | <font size="1">3.- Add 1 mL of RBC buffer and mix throughly by
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| - | vortex.</font><br />
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| - | <font size="1">4.- Centrifuge at 4,500 rpm during 30 s and pour
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| - | carefully so your pellet is not wasted.</font><br />
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| - | <font size="1">5.- Repeat steps 2 and 3 adding just 500 uL from
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| - | RBC buffer.</font><br />
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| - | <font size="1">6.- Once separated, add 300 uL of lysis buffer to
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| - | the pellet and mix by vortex.</font><br />
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| - | <font size="1">7.- Add 150 uL of protein precipitation solution.</font><br />
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| - | <font size="1">8.- Centrifuge at 11,000 rpm during 5 min; then,
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| - | transfer the supernatant to a new microtube.</font><br />
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| - | <font size="1">9.- Add 700 uL of cold isopropanol and mix by
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| - | inverting until you see a white formation within the tube.</font><br />
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| - | <font size="1">10.- Centrifuge at 11,000 rpm during 4 min and pour
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| - | carefully taking care of the pellet at the bottom.</font><br />
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| - | <font size="1">11.- Add 500 uL of cold ethanol 70% to wash and
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| - | then centrifuge at 11,000 rpm during 4 min.</font><br />
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| - | <font size="1">12.- Pour and let the ethanol evaporate at room
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| - | temperature.</font><br />
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| - | <font size="1"> 13.- Resuspend the DNA pellet with 12 uL of TE
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| - | buffer; incubate all night at 4 °C.</font><br />
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| - | </p>
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| - | </div>
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| - | <div>
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| - | <h1>Protocol 4</h1>
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| - | <p>Electrophoresis:<br />
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| - | <br />
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| - | <font size="1">1.- Prepare 1L of TAE 10X (48.4g of Tris base[ tris
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| - | (hidroxymethyl) amminomethane], 11.4 mL of glacial acetic acid
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| - | (17.4 M) and 3.7 g of EDTA then fill with deionized water to 1
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| - | L.</font><br />
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| - | <font size="1">2.- Prepare a 50 mL solution of 3% agarose using
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| - | TAE buffer (without using water).</font><br />
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| - | <font size="1">3.- Heat the gel in the microwave in intervals of
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| - | 30 s, 20 s and 10 s until the agarose is completely dissolved,
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| - | taking care that it does not boil so much.</font><br />
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| - | <font size="1">4.- Let cool down the gel (55-60°C) and pour in a
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| - | base with the comb already on its place to form the charging
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| - | wells.</font><br />
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| - | <font size="1">5.- Wait until the gel solidifies.</font><br />
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| - | <font size="1">6.- Remove the comb and place the base with the gel
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| - | inside the electrophoretic chamber.</font><br />
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| - | <font size="1">7.- Add TAE buffer until the gel is covered.</font><br />
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| - | <font size="1">8.- Take 5 uL of each sample from the PCR and 1 uL
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| - | of charging gel in a new PCR tube, mix well.</font><br />
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| - | <font size="1">9.- Charge the samples inside the wells of the gel,
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| - | adding a weight marcker.</font><br />
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| - | <font size="1">10.- Close the electrophoretic chamber and run at
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| - | 120 V during 30 min. Follow the displacement ofthe samples.</font><br />
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| - | <font size="1">11.- Take out the gel from the chamber very
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| - | carefully and place in the UV light to see the DNA strands.</font><br />
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| - | </p>
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| - | </div>
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| - | <div>
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| - | <h1>Protocol 5</h1>
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| - | <p>Competent cell transformation:<br />
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| - | <br />
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| - | 1.- Place 1 uL of the plasmid in a microtube, gently add 100 uL of
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| - | your competent cells. Take another tube and place only 100 uL of
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| - | competent cells, so you have a control group.<br />
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| - | 2.- Mix gently with a pipette and incubate during 30 min in ice.<br />
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| - | 3.- Ab ruptly place your cells in 42°C dry bath during 2 min and
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| - | replace in ice whwn finished.<br />
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| - | 4.- Add 900 uL of LB medium and incubate at 37 °C for 30 min.<br />
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| - | 5.- Inoculate with 200 uL of your transformed cells in LB-Amp.<br />
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| - | 6.- Incubate overnight and store at 4 °C of freeze in 15-25%
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| - | glycerol.<br />
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| - | </p>
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| - | </div>
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| - | <div> <img alt="" src='https://static.igem.org/mediawiki/2014hs/e/e4/LabArchives.jpg' width='400' height='300' />
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| - | <h1>Lab archives// Mar 12, 2014 </h1>
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| - | <p>In order to grow our bacteria, we inoculated with 100 uL of E.
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| - | Coli DH5-alpha in 50 mL of LB medium, and 100 uL of E. Coli TOP in
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| - | 50 mL of LB medium. We left both samples incubating overnight </p>
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| - | <h1>Lab archives// Mar 20, 2014 </h1>
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| - | <p>We inoculated E.Coli NEB10-beta in 50 ml of LB medium. </p>
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| - | </div>
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| - | <div>
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| - | <h1>Lab archives// Mar 27, 2014 </h1>
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| - | <p>We prepared the medium in which our bacteria will grow, for that
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| - | we dissolved solid LB with agarose, then we add it into 10 plates,
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| - | we waited a few minutes for it to solidify and finally we added
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| - | 500 uL of our antibiotic that is chloranphenicol to each of those
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| - | plates. </p>
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| - | <h1>Lab archives// Apr 01, 2014 </h1>
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| - | <p>we transformed luciferaze from DNA to cells by incerting them
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| - | with change of temperature </p>
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| - | <h1>Lab archives// Apr 02, 2014 </h1>
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| - | <p>We made a litter of agar and 500 L of growth medium, and we
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| - | sterilized them along with crystal media disperssion balls </p>
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| - | <h1>Lab archives// Apr 03, 2014 </h1>
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| - | <p>Today we prepared buffers for home made mini preps and did a cell
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| - | re culture. </p>
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| - | </div>
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| - | <div>
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| - | <h1><font size="1">Lab archives// Apr 07, 2014 </font> </h1>
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| - | <p> <font size="1">We added 700 mL of water in a beaker and then we
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| - | prepared 23 grams of powdered agar that we added slowly while we
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| - | moved the beaker. This in order to help the dissolve the powder.
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| - | After that, we added other 300 mL to the solution and finally we
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| - | passed it to an Erlenmeyer matrass. Also we prepared 500 mL of
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| - | growth medium. First, we measured and added 5 grams of tryptone,
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| - | 5 grams of yeast and 10 grams of sodium chloride. Those
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| - | ingredients were added slowly to a graduated cylinder with 350
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| - | mL of wáter while the flask was moved. After finishing the
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| - | addition we added other 150 mL of water. The solution was then
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| - | transfered to another Erlenmeyer flask. Both Erlenmeyer flasks
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| - | and a glass bottle with crystal media diperssion balls were
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| - | sterilized with temperature and pressure.</font> </p>
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| - | <h1><font size="1">Lab archives// Apr 02, 2014 </font> </h1>
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| - | <p><font size="1">300 ul of ampicillin at 100mg/ml chloramphenicol </font><br />
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| - | <font size="1"> 3 Plates with agar </font><br />
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| - | <font size="1">Promoters: BBa_K258005 (pO2), BBa_176500 (pFe)
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| - | BBa_J52008 (luc-3) </font> <br />
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| - | <font size="1">The plates were impregnated with ampicillin
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| - | promoting bacteria in the following manner </font><br />
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| - | <font size="1"> |pO2 | Plate | 150 ul Ampiciline | K258005 | </font><br />
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| - | <font size="1">
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| - | ------------------------------------------------------ </font><br />
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| - | <font size="1"> |pFe | Plate | 150 ul Ampiciline | 1765000 | </font><br />
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| - | <font size="1">
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| - | ------------------------------------------------------ </font><br />
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| - | <font size="1"> |Luc-3 | Plate | Chloranphenicol | J52008 | </font><br />
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| - | </p>
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| - | <p></p>
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| - | </div>
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| - | <div>
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| - | <h1><font size="1">Apr 08, 2014 </font></h1>
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| - | <p> <font size="1"> **/Today we realized the mini prep for the
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| - | oxygen promoter. /** </font><br />
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| - | <font size="1"> --500 ul of Ampiciline //this is for the oxygen
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| - | promoter </font><br />
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| - | <font size="1"> --50 ul of oxygen promoter </font><br />
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| - | <font size="1"> --45 ml of LB medium </font><br />
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| - | <font size="1"> --at room temperature 15 minutes at 1500 rpm in
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| - | the centrifuge. </font><br />
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| - | <font size="1"> --500 ul of chloramphenicol // this is for the red
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| - | liciferase </font><br />
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| - | <font size="1"> --50 ul of red luciferase </font><br />
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| - | <font size="1"> --45 ml of LB medium </font><br />
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| - | <font size="1"> --The new LB medium was prepared with //LB medium
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| - | </font><br />
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| - | <font size="1"> --10 g yeast </font><br />
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| - | <font size="1"> --5 g salt </font><br />
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| - | <font size="1"> --15 g Tryptone </font> <font size="1"> MINI
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| - | PREP First 400 ul of Genomic Lysis Buffer to the 40 ml overnight
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| - | [bacteriaand oxygen promoter] Then the mixture is transferred to
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| - | a Zyne-Spine Collumn in a Collection Tube. Centrifuge 10,000 rpm
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| - | /1 min Transfer the Zyno Spin Column to a new Collection Tube.
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| - | Add 200 micro liters of Pre Wash Buffer. Centrifuge 10,000 rpm
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| - | /1 min Add 500 uL of of g-DNA Wash buffer to the spin column.
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| - | Centrifuge 10,000 rpm /1 min Transfer the spin column to a clear
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| - | micro centrifuge tube. Add 50 uL of DNA Elution Buffer .
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| - | Incubate 2-5 minutes to at room temperature. Centrifuge at top
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| - | speed per 30 seconds. Then store at -80 °C </font></p>
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| - | </div>
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| - | <div>
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| - | <h1><font size="1"> Apr 22, 2014</font> </h1>
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| - | <p><font size="1"> We prepared 2 overnight cultures (45ml of LB
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| - | medium): GFP E0040 pSB1A2 with 100 uL of ampicillin. M-cherry
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| - | (RFP) BBa_J04450 pSB1C3 with 500 uL of chloramphenicol.</font></p>
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| - | <h1><font size="1"> Apr 24, 2014 </font></h1>
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| - | <p><font size="1"> 2 Transformations: Bacteria NEB 10 Beta, DH5alpha
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| - | 2 Mini Preps: GFP, RFP //Transformation and preps:--
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| - | Centrifugation at 2500 rpm for 15 min.--1ul DNA in 100 uL of
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| - | bacterial culture (luciferase). --Filtering GFP, RFP bacteria.
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| - | --Added 400 ml of Lysis Buffer Genomyc each crop by columns.
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| - | --Centrifugation at 10,000 * g for 1 min. --Last column to
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| - | another tube and DNA Prewash + 200 uL Buffer. --Centrifuged at
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| - | 10,000 * g for 1 min. --Add 500ul of DNA Wash Buffer,
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| - | centrifuged at 10,000 * g for 1 min. --Transfer to clean tube,
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| - | add 50ul of DNA Elution Buffer. --Incubate for 2-5 min at room
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| - | temperature, centrifuged at maximum speed for 30 sec.</font></p>
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| - | <h1><font size="1"> Apr 28, 2014 </font></h1>
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| - | <p><font size="1"> Today was prepared LB media. It also was done the
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| - | sterilization of 1 mL and 200 uL tips, water and ependorfs.
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| - | After the sterilization of the LB media was done, we cultivated
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| - | in it the bacteria carrying the oxygen promoter, RFP, and GFP.
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| - | Then we left the bacteria growing in the incubator at 37°C
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| - | overnight.</font></p>
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| - | </div>
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| - | <div>
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| - | <h1><font size="1"> May 05, 2014 </font></h1>
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| - | <p><font size="1"> Plating bacteria with GFP, RFP, promoter Oxygen
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| - | and iron promoter plasmids. Everyone in their respective
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| - | environments and specific antibiotics (ampicillin and
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| - | chloramphenicol). Tubes with bacteria was added and placed them
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| - | in LB medium incubator to grow more product if necessary.</font></p>
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| - | <h1><font size="1"> May 08, 2014 </font></h1>
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| - | <p><font size="1"> We did a centrifuge at 4800 rpm for 20 min for
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| - | the plasmids had been in overnight which had oxygen promoter,
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| - | GFP and RFP to rush after we separate the aqueous phase.</font></p>
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| - | <h1><font size="1"> May 19, 2014 </font></h1>
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| - | <p><font size="1"> We did inoculation of GFP, RFP and Oxigen Prmoter</font></p>
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| - | <h1><font size="1"> May 20, 2014 </font></h1>
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| - | <p><font size="1"> We started minipreps preparation of GFP, RFP and
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| - | Oxigen Promoter. We added 1.5 ml of culture to each Eppendorff
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| - | (4 for each promoter) and we centrifugated it at 13.2 rpm for 10
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| - | minutes, we did the same procedure 2 times. Also, we sterilized
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| - | two Erlenmeyer flasks and 40 Eppendorffs. </font></p>
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| - | </div>
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| - | <div>
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| - | <h1><font size="1"> May 20, 2014 //Mini prep preparation </font></h1>
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| - | <p><font size="1"> 1. We added 1.5 mL of the cultures to their
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| - | corresponding Eppendorffs and centrifugated them at 3400 rpm
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| - | during 10 minutes other three times.</font></p>
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| - | <p><font size="1">2. Once we had a pellet we discarded the
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| - | supernatant and we added 300 uL of TEG buffer (made of 100 mM of
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| - | Tris-HCl pH8, 2 mM EDTA and 20% glucose)</font></p>
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| - | <p><font size="1">3. We added 700 μL of NS solution (0.2 M NaOH, 1%
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| - | w/v SDs) and we mixed by inversion.</font></p>
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| - | <p><font size="1">4. We added 400 μL of 3 M sodium acetate pH 5.3,
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| - | and we mixed by inversion.</font></p>
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| - | <p><font size="1">5. We centrifugated the Eppendorffs at 3400 rpm
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| - | during 15 minutes.</font></p>
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| - | <p><font size="1">6. We transfered the supernatant to other
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| - | Eppendorffs and we added 50 uL of cold isopropanol and mixed by
| |
| - | inversion.</font></p>
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| - | <p><font size="1">7. We centrifugated the Eppendorffs at 3400 rpm
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| - | during 15 minutes.</font></p>
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| - | <p><font size="1">8. We discarded the supernatant and washed each
| |
| - | pellet with 70% ethanol and we let it dry for 15 minutes.</font></p>
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| - | <p><font size="1">9. We resuspended the pellet in 500 uL of TE
| |
| - | buffer and then we added 10 mg/mL RNAseA.</font></p>
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| - | <p><font size="1">11. We precipitated the DNA with 100 uL of ethanol
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| - | in each Eppendorff.</font></p>
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| - | <p><font size="1">12. We centrifugated for 1 minute at 3400 rpm.</font></p>
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| - | <p><font size="1">13. We got rid of the supernatant.</font></p>
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| - | <p><font size="1">14. Finally, we resuspended the pellet in 50 uL of
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| - | TE buffer</font></p>
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| - | </div>
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| - | <div> <img alt="" src='https://static.igem.org/mediawiki/2014hs/f/f4/1%281%29.jpg' width='400' height='300' />
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| - | <h1>UNAM procedures// Digestion</h1>
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| - | <p>|BSA | 1 uL | </p>
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| - | <p>|DNA | 2 uL | </p>
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| - | <p>|Buffer (2) | 1 uL |</p>
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| - | <p>|Buffer Xba I+Pstl | 3.3 uL |</p>
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| - | <p>|fe Xba I+Spef | 3.3 uL | </p>
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| - | <p>|H<sub>2</sub>0 | 5.4 uL | </p>
| |
| - | </div>
| |
| - | <div>
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| - | <h1><font size="1">Gel agarose</font></h1>
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| - | <p><font size="1"> 1 kb NEB Ampyciline -> Pentneryl </font></p>
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| - | <p><font size="1">| ---------------Bristol - Myers Squibb</font> </p>
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| - | <p><font size="1">| ---------------1 gram/ 3 mL</font> </p>
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| - | <p><font size="1">| ---------------inyectable</font></p>
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| - | <p><font size="1">|<sub>p</sub>O <sub>2</sub> | 3.3 uL</font> </p>
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| - | <p><font size="1">|1 kb marker 5.4 uL </font></p>
| |
| - | <h1><font size="1">Homemade Mini prep Preparation </font></h1>
| |
| - | <p><font size="1"> 1.- Overnight bacterial culture of 5 mL in
| |
| - | falkons of 50 mL </font></p>
| |
| - | <p><font size="1"> 2.- Centrifugate in an eppendor of 1.5 mL and
| |
| - | throw supernadant</font></p>
| |
| - | <p><font size="1"> 3.- Add 152 uL of Buffer 1 + RNAse (11
| |
| - | micro-liters /1 buffer micro-liter ) **Buffer 20% Glucouse </font></p>
| |
| - | <p><font size="1"> 4.- Mix via Buffer and add 150 uL of Buffer 2
| |
| - | *Wait 4-5 minutes*</font> </p>
| |
| - | <p><font size="1"> (1.5mL)Buffer 2 -> 880 uL H<sub>2</sub>O,20uL
| |
| - | NaOH 10 N </font></p>
| |
| - | <p><font size="1"> ---------------------------------------100 uL SDS
| |
| - | 10%</font> </p>
| |
| - | <p><font size="1"> 5.- Mix via inversion and add150 uL Buffer 3 </font>
| |
| - | </p>
| |
| - | <p><font size="1"> ------------------------------Sodium Acetate 3M
| |
| - | pH 5</font> </p>
| |
| - | </div>
| |
| - | <div>
| |
| - | <p><font size="1"> 6.-Centrifuge 5 minutes.</font> </p>
| |
| - | <p><font size="1"> 7.-Take the supernadant to antoher tube. Add 1 mL
| |
| - | ETOH Absolute </font></p>
| |
| - | <p><font size="1"> 8.-Centrifuge by 5 minutes and decant </font> </p>
| |
| - | <p><font size="1"> 9.-Wash with 500uL ETOH 70 %</font> </p>
| |
| - | <p><font size="1"> 10.-Centrifugate 1 minute </font></p>
| |
| - | <p><font size="1"> 11.-Decant and dry at 37 celsius centigrades (30
| |
| - | minutes)</font> </p>
| |
| - | <p><font size="1"> 12.-Resuspend in 15 uL of H<sub>2</sub>O </font></p>
| |
| - | <h1><font size="1">Ligation</font></h1>
| |
| - | <p><font size="1"> Inserto 7mL</font></p>
| |
| - | <p><font size="1"> Plasmid 3mL</font> </p>
| |
| - | <p><font size="1"> Buffer 10x 1.5mL </font> </p>
| |
| - | <p><font size="1"> Ligase 0.5 mL </font></p>
| |
| - | <p><font size="1"> H<sub>2</sub>O 3 mL </font> </p>
| |
| - | <p><font size="1"> Total = 15 ml </font></p>
| |
| - | </div>
| |
| - | <div>
| |
| - | <h1>E.Coli cells transformation. </h1>
| |
| - | <p>To transform E.Coli cells with an exogen DNA, 200 uL of competent
| |
| - | cells were taken they were mixed with 1-20 ug(Micrograms) of DNA
| |
| - | that was incubated on ice for 30 minutes. This makes posible the
| |
| - | absorbtion of the exogen DNA to the cells surface, after that, the
| |
| - | cells are incubated at 42Celsius Degrees during 90 seconds, for
| |
| - | then incubate them during 1 minute again in ice. THis termal shock
| |
| - | makes posible the entrance of the plasmid to the inside of the
| |
| - | cell. 600 uL of LB medium were added and then incubated during
| |
| - | 60-90 minutes at 37 Celisus Degres, to do posible the segregation
| |
| - | of the exogenic plasmid during the celular division (3 to 5
| |
| - | generations). Finally, Alicuote of 100-200 uL were cultivated by
| |
| - | dimention in the LB agar medium, which ere incubated at 37 Celsius
| |
| - | Degrees. After 15-20 hours of incuvation colonies could be
| |
| - | observed</p>
| |
| - | </div>
| |
| - | <div>
| |
| - | <h1>Ligations</h1>
| |
| - | <p>1# Grow h mL in overnight</p>
| |
| - | <p>2# Do minipreps</p>
| |
| - | <p>3# EcoRI and PstI restrictions.</p>
| |
| - | <p> --for Fe EcoRIand SpeI</p>
| |
| - | <p>O<sub>2</sub>+ GFP ---1,2,3 - 931</p>
| |
| - | <p>Fe+GTP ---4,5,6 -1,828</p>
| |
| - | <p>O<sub>2</sub>+ RFP ---7,8,9,10,11,12,13 -890</p>
| |
| - | <p>Fe+RFP ---14,15,16,17,18,19 -1,787</p>
| |
| - | <br />
| |
| - | <p>GFP -> 761 bp EcoRI Po+I // 728 bp XbaI SpeI</p>
| |
| - | <p>RFP -> 714 bp Xba - SpeI // 747 bp EcoR - Ps+I</p>
| |
| - | <br />
| |
| - | <p>pFe -> 1,067 bp EcoRI - SpeI </p>
| |
| - | <p><sub>p</sub>O<sub>2</sub> -> 178 bp EcoRi SpeI // 145 bp Xba -
| |
| - | Ps+I </p>
| |
| - | </div>
| |
| - | <div>
| |
| - | <p>O<sub>2</sub> + GFP 1/1 - 1/2 - 3/3</p>
| |
| - | <p>FO <sub>2</sub> + GFP 4/1 - 5/2 - 6/3</p>
| |
| - | <p>O<sub>2</sub> + RFP 7/1 - 8/2 - 9/3 - 10/4 - 11/5 - 12/6 - 12/13
| |
| - | </p>
| |
| - | <p>Fe + RFP 13/1 - 14/2 - 15*16/3 - 17/4 - 18/5 -19/6</p>
| |
| - | <h1>Digestion</h1>
| |
| - | <p>2 --- BSA --- 1uL</p>
| |
| - | <p>4 --- DNA --- 2uL</p>
| |
| - | <p>3 --- (M) Buffer // Enz (0.3 uL each one)--- 1uL</p>
| |
| - | <p>5 --- EcoRI --- 0.3 uL</p>
| |
| - | <p> --- PstI --- 0.3 uL</p>
| |
| - | <p>1 --- H<sub>2</sub>O --- 5.4 uL</p>
| |
| - | <p> Total == 10 uL rxn</p>
| |
| - | <br />
| |
| - | <p> GFP 761 bp</p>
| |
| - | <p> RFP 742 bp</p>
| |
| - | </div>
| |
| - | <div>
| |
| - | <p> pFe 1,067 bp </p>
| |
| - | <p> pO<sub>2</sub> -15.5 bp</p>
| |
| - | <br />
| |
| - | <p> pO<sub>2</sub> + GFP 931 bp</p>
| |
| - | <p> pFe + GFP 1,828 bp</p>
| |
| - | <p> pO<sub>2</sub>+ RFP 890 bp</p>
| |
| - | <p> pFe + RFP 1,787 bp</p>
| |
| - | <br />
| |
| - | <p> Gel 1 = 8/9/10/Lodder 1kb/11/12/13</p>
| |
| - | <p> Gel 2 = 15/16/17/Lodder 116/18/19</p>
| |
| - | <p> Gel 3 = 1/2/3/4/Lodder 1kb/5/6/7</p>
| |
| - | </div>
| |
| - | </div>
| |
| - | </div>
| |
| - | </div>
| |
| - | <div> <span class="reference"> </span> </div>
| |
| - | <!-- The JavaScript -->
| |
| - | <script type="text/javascript">
| |
| - | $(function() {
| |
| - | var $mybook = $('#mybook');
| |
| - | var $bttn_next = $('#next_page_button');
| |
| - | var $bttn_prev = $('#prev_page_button');
| |
| - | var $loading = $('#loading');
| |
| - | var $mybook_images = $mybook.find('img');
| |
| - | var cnt_images = $mybook_images.length;
| |
| - | var loaded = 0;
| |
| - | //preload all the images in the book,
| |
| - | //and then call the booklet plugin
| |
| - |
| |
| - | $mybook_images.each(function(){
| |
| - | var $img = $(this);
| |
| - | var source = $img.attr('src');
| |
| - | $('<img/>').load(function(){
| |
| - | ++loaded;
| |
| - | if(loaded == cnt_images){
| |
| - | $loading.hide();
| |
| - | $bttn_next.show();
| |
| - | $bttn_prev.show();
| |
| - | $mybook.show().booklet({
| |
| - | name: null, // name of the booklet to display in the document title bar
| |
| - | width: 800, // container width
| |
| - | height: 500, // container height
| |
| - | speed: 600, // speed of the transition between pages
| |
| - | direction: 'LTR', // direction of the overall content organization, default LTR, left to right, can be RTL for languages which read right to left
| |
| - | startingPage: 0, // index of the first page to be displayed
| |
| - | easing: 'easeInOutQuad', // easing method for complete transition
| |
| - | easeIn: 'easeInQuad', // easing method for first half of transition
| |
| - | easeOut: 'easeOutQuad', // easing method for second half of transition
| |
| - |
| |
| - | closed: true, // start with the book "closed", will add empty pages to beginning and end of book
| |
| - | closedFrontTitle: null, // used with "closed", "menu" and "pageSelector", determines title of blank starting page
| |
| - | closedFrontChapter: null, // used with "closed", "menu" and "chapterSelector", determines chapter name of blank starting page
| |
| - | closedBackTitle: null, // used with "closed", "menu" and "pageSelector", determines chapter name of blank ending page
| |
| - | closedBackChapter: null, // used with "closed", "menu" and "chapterSelector", determines chapter name of blank ending page
| |
| - | covers: false, // used with "closed", makes first and last pages into covers, without page numbers (if enabled)
| |
| - |
| |
| - | pagePadding: 10, // padding for each page wrapper
| |
| - | pageNumbers: true, // display page numbers on each page
| |
| - |
| |
| - | hovers: false, // enables preview pageturn hover animation, shows a small preview of previous or next page on hover
| |
| - | overlays: false, // enables navigation using a page sized overlay, when enabled links inside the content will not be clickable
| |
| - | tabs: false, // adds tabs along the top of the pages
| |
| - | tabWidth: 60, // set the width of the tabs
| |
| - | tabHeight: 20, // set the height of the tabs
| |
| - | arrows: false, // adds arrows overlayed over the book edges
| |
| - | cursor: 'pointer', // cursor css setting for side bar areas
| |
| - |
| |
| - | hash: false, // enables navigation using a hash string, ex: #/page/1 for page 1, will affect all booklets with 'hash' enabled
| |
| - | keyboard: true, // enables navigation with arrow keys (left: previous, right: next)
| |
| - | next: $bttn_next, // selector for element to use as click trigger for next page
| |
| - | prev: $bttn_prev, // selector for element to use as click trigger for previous page
| |
| - |
| |
| - | menu: null, // selector for element to use as the menu area, required for 'pageSelector'
| |
| - | pageSelector: false, // enables navigation with a dropdown menu of pages, requires 'menu'
| |
| - | chapterSelector: false, // enables navigation with a dropdown menu of chapters, determined by the "rel" attribute, requires 'menu'
| |
| - |
| |
| - | shadows: true, // display shadows on page animations
| |
| - | shadowTopFwdWidth: 166, // shadow width for top forward anim
| |
| - | shadowTopBackWidth: 166, // shadow width for top back anim
| |
| - | shadowBtmWidth: 50, // shadow width for bottom shadow
| |
| - |
| |
| - | before: function(){}, // callback invoked before each page turn animation
| |
| - | after: function(){} // callback invoked after each page turn animation
| |
| - | });
| |
| - | Cufon.refresh();
| |
| - | }
| |
| - | }).attr('src',source);
| |
| - | });
| |
| - |
| |
| - | });
| |
| - | </script>
| |
| - | </body>
| |
| - | </html>
| |
"
