Team:MIT mmethods

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Bacterial Construction Protocol
Bacterial Experimental Protocol
Phage western blot
Mammalian Protocol
Microfluidic stress

Materials

mammalian methods

The Mammalian team focused on creating a pressure-sensitive promoter and creating a standard protocol for mammalian genes.

1 HTD Preparation Protocol
    1.1 PDMS Mixture Preparation
    1.2 PDMS Pouring
    1.3 PDMS Baking
    1.4 PDMS-Device punching and Bonding
    1.5 PDMS Device Bonding
    1.6 PDL coating
    1.7 Collagen filling
    1.8 Cell Seeding
2 Protocol for Deflection Experiments
    2.1 Tubing Setup Details
    2.2 Adding Medium to Channels
    2.3 Connecting device to pressure valve
    2.4 Microcontroller details

HTD Preparation Protocols (adapted from Yannis, Alisha)

PDMS Mixture Preparation
Material
  1. PDMS Body liquid (10 parts)
  2. PDMS curing agent (1 part)
  3. Negative pattern wafer
  4. Microscope slides (1mm thick)

Procedure
  1. Pour body liquid in a plastic cup while measuring its weight (Make sure to cover the balance with tissue)
  2. Reset scale, pour curing agent while measuring its weight (in 1:10 ratio to body liquid)
  3. Stir the mixture
  4. Remove the bubbles in the vacuum chamber
        a. Turn on vacuum and close the valve for sustaining vacuum (Make sure to cover the chamber with tissue)
    Note: Lift container to be surethe vacuum is on.
        b. De-gas for 20 minutes
    Note: Open/close valve quickly to get rid of bubbles faster. When finished, close vacuum; open valve slowly.