Team:USMA-West Point/Notebook

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USMA-West_Point

<!DOCTYPE html> Notebook - iGem 01

usMA-WEST POINT


Experiments



Day to Day activity inside the lab



January 2017



Transformation of Bacteria Cells



  1. OR vectors were purchased from OriGene and transfected into XL-10 Gold Ultracompetent cells by heat shocking at 42 degrees Celsius
  2. Cells were cultured in LB broth to amplify vectors
  3. DNA gels were run with specific restriction enzymes to determine multiple cloning sites
  4. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, 2LT Dillon Macky, 2LT Ishmael Raheem, CDT Jason Hug, CDT Elizabeth Huuki, CDT Channah Mills, CDT Matthew McDonough, Isaac Dabkowski


February 2017



Western Blotting



  1. HT-22 cells were lysed in RIPA buffer with phosphate and protease inhibitors

  2. Protein concentration was measured for each OR

  3. Proteins were prepared with Laemmle sample buffer before loading into 4% Bis-Tris gels to measure protein purity and size

  4. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, 2LT Dillon Macky, 2LT Ishmael Raheem, CDT Jason Hug, CDT Elizabeth Huuki, CDT Channah Mills, CDT Matthew McDonough, Isaac Dabkowski​


March 2017



Confocal Microscopy



  1. HT-22 cells were fixed with 4% paraformaldehyde and immunohistochemistry with antibodies responsible for individual OR binding followed by secondary antibodies for visualization

  2. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, 2LT Dillon Macky, 2LT Ishmael Raheem, CDT Jason Hug, CDT Elizabeth Huuki, CDT Channah Mills, CDT Matthew McDonough, Isaac Dabkowski


April 2017



Action Potential Measurements



  1. Nucleofected HT-22 cells were stimulated with individual odorants

  2. The neurons generated action potentials that were measured via a microelectrode array

  3. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, 2LT Dillon Macky, 2LT Ishmael Raheem, CDT Jason Hug, CDT Elizabeth Huuki, CDT Channah Mills, CDT Matthew McDonough, Isaac Dabkowski


May 2017



USMA Projects Day



  1. Presented research at United States Military Academy Projects Day, a day where students present projects concerning research in various fields

  2. Reports put together for eNOSE

  3. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, 2LT Dillon Macky, 2LT Ishmael Raheem, CDT Jason Hug, CDT Elizabeth Huuki, CDT Channah Mills, CDT Matthew McDonough, Isaac Dabkowski


June 2017



Further Trials and Biosensor Design



  1. Cells amplified

  2. Focused on designing a biosensor

    • Bioreactor
    • Microelectrode array (MEA)
    • Perfusion system
    • Signal Processor
  3. Participants
    1. Dr. Kirk Pirlo, Dr. Joel Gaston, Winston Ou, Rayonna Gordon, CDT Angela Marsh, CDT Kanak Pal, CDT Payton Boylston, CDT Matthew McDonough


July 2017



Finalizing Design



  1. Finalized biosensor design as necessary parts were delivered

  2. Prepared for trials with updated biosensor
  3. Additional transformations
  4. Participants
    1. Dr. Kirk Pirlo, Dr. Joel Gaston, Winston Ou, Rayonna Gordon, Dr. Alex Mitropoulos


August 2017



Trials with Updated Device



  1. Ran successful trials with updated biosensor

  2. Collected more data for device

  3. Participants
    1. Dr. Kirk Pirlo, Dr. Joel Gaston, Winston Ou, Rayonna Gordon, Dr. Alex Mitropoulos


September 2017



Interlab Protocol and iGEM Plasmid



  1. Completed Interlab protocol and collected data

  2. Developed iGEM plasmid by transfection and amplification

  3. Ran gel of plasmids to evaluate

  4. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, Dr. John Cave, CDT Jason Hug, CDT Zachary Andersen, CDT Kylor Keisewetter, CDT Alana Appel, CDT Elizabeth Huuki​, CDT Channah Mills, CDT Matthew McDonough, CDT James Pruneski


October 2017



Preparation for iGEM



  1. Finalized results

  2. Design and develop parts

  3. Prepared iGEM wiki and presentation

  4. Participants
    1. Dr. Alexander Mitropoulos, Dr. Ken Wickiser, Dr. Kamil Woronowicz, Dr. John Cave, CDT Jason Hug, CDT Zachary Andersen, CDT Kylor Keisewetter, CDT Alana Appel, CDT Elizabeth Huuki​, CDT Channah Mills, CDT Matthew McDonough, CDT James Pruneski


DEPARTMENT OF CHEMISTRY AND LIFE SCIENCE



BARTLETT HALL, ROOM 400, WEST POINT, NY 10996, USA


TEL: 845.938.3915