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| - | {{:Team:UC-SantaCruz_Template}}
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| - | 2013 UCSC IGEM TEAM Polar Tags Notebook.
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| - | Index-
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| - | Topic Page #
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| - | Groups Goals
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| - | The goal of the Polar Tag Group is to
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| - | General PCR Protocol
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| - | General Agarose Gel Protocol
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| - | General Chemicompetent Cell Transformation Protocol
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| - | LB Media Recipe
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| - | Colony Pick Protocol
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| - | Electroporation Protocol
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| - | DNA MW marker reference
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| - | [[File:Untitled 1.png|200px|left]]
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| - | Reference Sequences
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| - | References
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| - | Lab Notebook 8/6/13
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| - | Lab Notebook 8/7/13
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| - | Lab Notebook 8/8/13
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| - | Lab Notebook 8/9/13
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| - | Lab Notebook 8/13/13
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| - | Lab Notebook 8/15/13
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| - | Lab Notebook 8/16/13
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| - | Lab Notebook 8/19/13
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| - | Lab Notebook 8/20/13
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| - | Lab Notebook 8/21/13
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| - | Lab Notebook 8/22/13
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| - | Lab Notebook 8/25/13
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| - | Lab Notebook 8/26/13
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| - | Lab Notebook 8/27/13
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| - | Lab Notebook 8/28/13
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| - | Lab Notebook 8/29/13
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| - | Lab Notebook 8/30/13
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| - | Lab Notebook 8/31/13
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| - | Lab Notebook 9/1/13
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| - | Lab Notebook 9/2/13
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| - | Lab Notebook 9/3/13
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| - | Lab Notebook 9/4-5/13
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| - | Lab Notebook 9/6/13
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| - | Lab Notebook 9/7/13
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| - | Making Top10 Chemicompetent Cells:
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| - | 1) Grow Top10 SOB overnight between the temperatures 20°C and 33°C.
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| - | 2) Measure the OD and bring the OD back to 0.1.
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| - | 3) Add 2.5 ml of Top10 SOB to SOB.
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| - | 4) Incubate at 31°C for one hour and 20 minutes or until the OD is 0.6.
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| - | 5) Create 5 ml of 1X Washing buffer: 2.5 ml of Dilution buffer and 2.5 ml of Wash buffer (2X)
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| - | 6) Create 5 ml of 1X Competent buffer: 2.5 ml Dilution buffer and 2.5 ml of Competent buffer (2X)
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| - | 7) Incubate the culture on ice for ten minutes and then pellet cells at 2,500 xg for ten minutes at 0-4°C.
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| - | 8) Remove supernatant. Gently resuspend cells in 5 ml of ice cold 1X Wash buffer. Re-pellet as in step 7.
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| - | 9) Remove supernatant completely. Gently resuspend cells in 5 ml of ice cold 1X Competent buffer.
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| - | 10) Aliquot 100-200 ul. Flash freeze.
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| - | 11) Store in -80°C freezer.
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| - | Making LB+Kanamycin and LB+Gentamicin Agar Plates:
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| - | To make a 500 ml batch:
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| - | - 475 ml of MilliQ water
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| - | - 5 g of Tryptone
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| - | - 5 g of NaCl
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| - | - 2.5 g of yeast Extract
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| - | - 7.5 g of Agar
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| - | We made x2 500 ml batches.
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| - | Combine the reagents and shake until the solutes have dissolved. Adjust the ph to 7.0. Sterilize by autoclaving for 20 minutes on liquid cycle.
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| - | Take out of the autoclave and let the solution cool. Place in a water bath to prevent solid formation.
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| - | Add 1.25 ml of gentamicin antibiotic to one of the 500 ml batches and 500 ul of kanamycin to the other batch.
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| - | Pour into petri dishes about half way. Let agar solidify. Place in the refrigerator.
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| - | Lab Notebook 9/8/13
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| - | Lab Notebook 9/10/13
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| - | Lab Notebook 9/11/13
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| - | Lab Notebook 9/12/13
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| - | Lab Notebook 9/13/13
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| - | Lab Notebook 9/14/13
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| - | Lab Notebook 9/15/13
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| - | Lab Notebook 9/16/13
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| - | Lab Notebook 9/17/13
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| - | Lab Notebook 9/18/13
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| - | Lab Notebook 9/19/13
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| - | Lab Notebook 9/20/13
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