Team:Northwestern/Protocol

From 2010.igem.org

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===[[LIVE/DEAD® BacLight - Bacterial Viability Kit]]===
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Revision as of 23:47, 9 October 2010


Tech Institute
Home Brainstorm Team Acknowledgements Project Human Practices Parts Notebook Calendar Protocol Safety Links References Media Contact


DNA

Prepping DNA from the kit plates

Quikchange (from primers to colonies!)

Kit to Stock Plasmid

Ethanol Precipitation

New Part Design(PCR)

Bacterial Work

Transformation

O/N Culture

Preparation of Competent Cells

LB Media

Preparing Plates

Glycerol Stocks

Assembly

Restriction Enzyme Digests

Plasmid Construction

3A Assembly

Ligations

Mini Prep

Microscopy

Confocal Microscopy

Fluorescence Microscopy

Reagents

Reagents

Cell Staining

Rhodamine-Conjugated Chitin Probe

Methanol Fixation

  1. Rinse slide with ethanol and flame
  2. Rinse slide with PBS and dry with KimWipe
  3. Apply cell sample to microscope slide and let air dry
  4. Submerge in -20C absolute methanol for 5-10 min
  5. Wash 3 times with 1X TBS (submerge in 1X TBS for 5 minutes per wash)
    • Do not let cells dry for the rest of the procedure


LIVE/DEAD® BacLight - Bacterial Viability Kit

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