Team:Minnesota/Notebook

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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 [2] into TOP10 cells
  8. The transformants are plated then screened[3] using a florescent camera
  9. Once the colonies have been screened, positive colonies have their plasmids isolatedisolated.
  10. The plasmids are then [4] to ensure the correct sequence was obtained.



  11. 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
                [http://2009.igem.org/wiki/index.php?title=Minnesota/1_June_2008&action=edit 1]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/2_June_2008&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Minnesota/3_June_2008&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Minnesota/4_June_2008&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Minnesota/5_June_2008&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Minnesota/6_June_2008&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Minnesota/7_June_2008&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Minnesota/8_June_2008&action=edit 8]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/9_June_2008&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Minnesota/10_June_2008&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Minnesota/11_June_2008&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Minnesota/12_June_2008&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Minnesota/13_June_2008&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Minnesota/14_June_2008&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Minnesota/15_June_2008&action=edit 15]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/16_June_2008&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Minnesota/17_June_2008&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Minnesota/18_June_2008&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Minnesota/19_June_2008&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Minnesota/20_June_2008&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Minnesota/21_June_2008&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Minnesota/22_June_2008&action=edit 22]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/23_June_2008&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Minnesota/24_June_2008&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Minnesota/25_June_2008&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Minnesota/26_June_2008&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Minnesota/27_June_2008&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Minnesota/28_June_2008&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Minnesota/29_June_2008&action=edit 29]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/30_June_2008&action=edit 30]
    July
    MTWTFSS
      [http://2009.igem.org/wiki/index.php?title=Minnesota/1_July_2008&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Minnesota/2_July_2008&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Minnesota/3_July_2008&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Minnesota/4_July_2008&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Minnesota/5_July_2008&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Minnesota/6_July_2008&action=edit 6]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/7_July_2008&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Minnesota/8_July_2008&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Minnesota/9_July_2008&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Minnesota/10_July_2008&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Minnesota/11_July_2008&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Minnesota/12_July_2008&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Minnesota/13_July_2008&action=edit 13]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/14_July_2008&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Minnesota/15_July_2008&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Minnesota/16_July_2008&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Minnesota/17_July_2008&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Minnesota/18_July_2008&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Minnesota/19_July_2008&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Minnesota/20_July_2008&action=edit 20]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/21_July_2008&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Minnesota/22_July_2008&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Minnesota/23_July_2008&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Minnesota/24_July_2008&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Minnesota/25_July_2008&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Minnesota/26_July_2008&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Minnesota/27_July_2008&action=edit 27]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/28_July_2008&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Minnesota/29_July_2008&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Minnesota/30_July_2008&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Minnesota/31_July_2008&action=edit 31]
    August
    MTWTFSS
            [http://2009.igem.org/wiki/index.php?title=Minnesota/1_August_2008&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Minnesota/2_August_2008&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Minnesota/3_August_2008&action=edit 3]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/4_August_2008&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Minnesota/5_August_2008&action=edit 5] [http://2009.igem.org/Minnesota/6_August_2008 6] [http://2009.igem.org/wiki/index.php?title=Minnesota/7_August_2008&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Minnesota/8_August_2008&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Minnesota/9_August_2008&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Minnesota/10_August_2008&action=edit 10]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/11_August_2008&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Minnesota/12_August_2008&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Minnesota/13_August_2008&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Minnesota/14_August_2008&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Minnesota/15_August_2008&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Minnesota/16_August_2008&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Minnesota/17_August_2008&action=edit 17]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/18_August_2008&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Minnesota/19_August_2008&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Minnesota/20_August_2008&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Minnesota/21_August_2008&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Minnesota/22_August_2008&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Minnesota/23_August_2008&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Minnesota/24_August_2008&action=edit 24]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/25_August_2008&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Minnesota/26_August_2008&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Minnesota/27_August_2008&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Minnesota/28_August_2008&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Minnesota/29_August_2008&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Minnesota/30_August_2008&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Minnesota/31_August_2008&action=edit 31]
    September
    MTWTFSS
    [http://2009.igem.org/wiki/index.php?title=Minnesota/1_September_2008&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Minnesota/2_September_2008&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Minnesota/3_September_2008&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Minnesota/4_September_2008&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Minnesota/5_September_2008&action=edit 5] [http://2009.igem.org/wiki/index.php?title=Minnesota/6_September_2008&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Minnesota/7_September_2008&action=edit 7]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/8_September_2008&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Minnesota/9_September_2008&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Minnesota/10_September_2008&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Minnesota/11_September_2008&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Minnesota/12_September_2008&action=edit 12] [http://2009.igem.org/wiki/index.php?title=Minnesota/13_September_2008&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Minnesota/14_September_2008&action=edit 14]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/15_September_2008&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Minnesota/16_September_2008&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Minnesota/17_September_2008&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Minnesota/18_September_2008&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Minnesota/19_September_2008&action=edit 19] [http://2009.igem.org/wiki/index.php?title=Minnesota/20_September_2008&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Minnesota/21_September_2008&action=edit 21]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/22_September_2008&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Minnesota/23_September_2008&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Minnesota/24_September_2008&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Minnesota/25_September_2008&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Minnesota/26_September_2008&action=edit 26] [http://2009.igem.org/wiki/index.php?title=Minnesota/27_September_2008&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Minnesota/28_September_2008&action=edit 28]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/29_September_2008&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Minnesota/30_September_2008&action=edit 30]
    October
    MTWTFSS
        [http://2009.igem.org/wiki/index.php?title=Minnesota/1_October_2008&action=edit 1] [http://2009.igem.org/wiki/index.php?title=Minnesota/2_October_2008&action=edit 2] [http://2009.igem.org/wiki/index.php?title=Minnesota/3_October_2008&action=edit 3] [http://2009.igem.org/wiki/index.php?title=Minnesota/4_October_2008&action=edit 4] [http://2009.igem.org/wiki/index.php?title=Minnesota/5_October_2008&action=edit 5]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/6_October_2008&action=edit 6] [http://2009.igem.org/wiki/index.php?title=Minnesota/7_October_2008&action=edit 7] [http://2009.igem.org/wiki/index.php?title=Minnesota/8_October_2008&action=edit 8] [http://2009.igem.org/wiki/index.php?title=Minnesota/9_October_2008&action=edit 9] [http://2009.igem.org/wiki/index.php?title=Minnesota/10_October_2008&action=edit 10] [http://2009.igem.org/wiki/index.php?title=Minnesota/11_October_2008&action=edit 11] [http://2009.igem.org/wiki/index.php?title=Minnesota/12_October_2008&action=edit 12]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/13_October_2008&action=edit 13] [http://2009.igem.org/wiki/index.php?title=Minnesota/14_October_2008&action=edit 14] [http://2009.igem.org/wiki/index.php?title=Minnesota/15_October_2008&action=edit 15] [http://2009.igem.org/wiki/index.php?title=Minnesota/16_October_2008&action=edit 16] [http://2009.igem.org/wiki/index.php?title=Minnesota/17_October_2008&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Minnesota/18_October_2008&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Minnesota/19_October_2008&action=edit 19]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/20_October_2008&action=edit 20] [http://2009.igem.org/wiki/index.php?title=Minnesota/21_October_2008&action=edit 21] [http://2009.igem.org/wiki/index.php?title=Minnesota/22_October_2008&action=edit 22] [http://2009.igem.org/wiki/index.php?title=Minnesota/23_October_2008&action=edit 23] [http://2009.igem.org/wiki/index.php?title=Minnesota/24_October_2008&action=edit 24] [http://2009.igem.org/wiki/index.php?title=Minnesota/25_October_2008&action=edit 25] [http://2009.igem.org/wiki/index.php?title=Minnesota/26_October_2008&action=edit 26]
    [http://2009.igem.org/wiki/index.php?title=Minnesota/27_October_2008&action=edit 27] [http://2009.igem.org/wiki/index.php?title=Minnesota/28_October_2008&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Minnesota/29_October_2008&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Minnesota/30_October_2008&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Minnesota/31_October_2008&action=edit 31]
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