Team:Northwestern/Protocol

DNA

Prepping DNA from the kit plates

Team:Northwestern/Protocol/Prepping DNA from the kit plates

Team:Northwestern/Protocol/Quikchange (from primers to colonies!)

Team:Northwestern/Protocol/Kit to Stock Plasmid

Team:Northwestern/Protocol/Ethanol Precipitation

Team:Northwestern/Protocol/New Part Design(PCR)

=Bacterial Work

Team:Northwestern/Protocol/Transformation

Team:Northwestern/Protocol/O/N Culture===

Team:Northwestern/Protocol/Preparation of Competent Cells

Team:Northwestern/Protocol/LB Media

Team:Northwestern/Protocol/Preparing Plates

Team:Northwestern/Protocol/Glycerol Stocks

Assembly

Team:Northwestern/Protocol/Restriction Enzyme Digests Team:Northwestern/Protocol/Plasmid Construction Team:Northwestern/Protocol/3A Assembly Team:Northwestern/Protocol/Ligations Team:Northwestern/Protocol/Mini Prep

Microscopy

Team:Northwestern/Protocol/Confocal Microscopy Team:Northwestern/Protocol/Fluorescence Microscopy

Reagents

Team:Northwestern/Protocol/Reagents

Cell Staining

Team:Northwestern/Protocol/Rhodamine-Conjugated Chitin Probe

Team:Northwestern/Protocol/Methanol Fixation

Methanol Fixation

Team:Northwestern/Protocol/LIVE/DEAD® BacLight - Bacterial Viability Kit

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