Team:Minnesota/Notebook

From 2009.igem.org

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<li>Combine [http://2009.igem.org/wiki/images/3/39/Soeing_PCR.pdf| Soeing PCR] reagents with primers and place in a thermocycler.</li>
<li>Combine [http://2009.igem.org/wiki/images/3/39/Soeing_PCR.pdf| Soeing PCR] reagents with primers and place in a thermocycler.</li>
<li>Amplify the insert using [http://2009.igem.org/wiki/images/3/39/PCR.pdf| PCR]</li>
<li>Amplify the insert using [http://2009.igem.org/wiki/images/3/39/PCR.pdf| PCR]</li>
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<li>Purify the PCR product using QIAquick PCR purification[http://2009.igem.org/wiki/images/3/39/PCR.pdf| PCR]</li>
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<li>Purify the PCR product using [http://2009.igem.org/wiki/images/3/39/DNA_Purification.pdf|QIAquick PCR purification]</li>
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<li></li>
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<li>Ligate[http://2009.igem.org/wiki/images/3/39/Ligation_Reaction.pdf|Ligation] the insert into pGLOTopo</li>
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<li></li>
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<li>The plasmid is transformed[http://2009.igem.org/wiki/images/3/39/Transformation_of_Chemically_Competent_Cells.pdf|transformed] into TOP10 cells</li>
<li></li>
<li></li>
<li></li>
<li></li>

Revision as of 18:44, 9 August 2009

Mnlogo.jpg
Home The Team The Project SynBioSS Designer Modeling Experimental Competition Requirements

Contents

Standard Protocols We Used in the Wet Lab

These link to pdf files:

Bacteria Culture Protocols

Transformation of Chemically Competent Cells

Plasmid Prep from Cultures

DNA Quantification

Polymerase Chain Reaction (PCR)

Restriction Digest

Vector Dephosphorylation

DNA Fragment Ligation

DNA Purification

Sequencing

Preparing Competent Cells

SOEing PCR

Ligation

Screening

Sample Collection

Procedure

  1. Prepare Competent Cells for TOP10 and DH5αPro
  2. Design the primers so that the proper mutations exist in the palindromic sequence of the Tet operator site. Design for each construct: TNN, TTN, and TTL.
  3. Combine Soeing PCR reagents with primers and place in a thermocycler.
  4. Amplify the insert using PCR
  5. Purify the PCR product using PCR purification
  6. Ligate[1] the insert into pGLOTopo
  7. The plasmid is transformed[2] into TOP10 cells


  8. Our Google Calendar

    This calendar contains a day-by-day catalog of what we did in the wet lab for our project and parts characterization. The calendar for computational work can be found below and on the Modeling page. Please click on each event to see a detailed description of what we did.



    Notebook




    June
    MTWTFSS
                1
    2 3 4 5 6 7 8
    9 10 11 12 13 14 15
    16 17 18 19 20 21 22
    23 24 25 26 27 28 29
    30
    July
    MTWTFSS
      1 2 3 4 5 6
    7 8 9 10 11 12 13
    14 15 16 17 18 19 20
    21 22 23 24 25 26 27
    28 29 30 31
    August
    MTWTFSS
            1 2 3
    4 5 6 7 8 9 10
    11 12 13 14 15 16 17
    18 19 20 21 22 23 24
    25 26 27 28 29 30 31
    September
    MTWTFSS
    1 2 3 4 5 6 7
    8 9 10 11 12 13 14
    15 16 17 18 19 20 21
    22 23 24 25 26 27 28
    29 30
    October
    MTWTFSS
        1 2 3 4 5
    6 7 8 9 10 11 12
    13 14 15 16 17 18 19
    20 21 22 23 24 25 26
    27 28 29 30 31