TISSUE CULTURE:
Best Practices
Preparing media
Splitting cells & seeding plates
Transfection (Lipofectamine LTX)
Transfection (Lipofectamine 2000)
Transfection (Lipofectamine 3000)
FACS Cell Prep
siRNA Transfection
.
Flow Cytometry with primary and secondary antibodies for membrane staining
Flow Cytometry recommended controls (from abcam)
Immunostaining for Fluorescent Microscopy
Immunostaining for Flow Cytometry
Gelatin Treatment for Glass Plates
Beta Amyloid Streptavidin Staining Protocol for Cytometry
Beta Amyloid Streptavidin Staining Protocol for Live Microscopy
Attaching B-Cells using Poly-L-Lysine
Freezing and Thawing HEK293s
ASSEMBLY, CLONING & ANALYSIS:
Gel (preparation, running & extraction)
Golden Gate
LR (Gateway)
Making liquid culture
Midiprep
Miniprep
PCR
Resuspending PCR primers
Restriction Digest
Transformation
BP
Annealing and Kinasing Oligos
PCR Purification
Designing and Ordering Primers
Site Directed Mutagenesis
MISC. LABORATORY PROCEDURES:
Making a Glycerol Stock
Making SOC
Preparing GGDonr plates
Preparing LB Agar plates
Western Blot
Oligomerizing Beta Amyloid
Autoclave
NON-LABORATORY PROCEDURES:
Checking T
m
of primers and other primer design tips
Locating a BsaI-Q cut site
Mutating DNA sequence to remove cut site
Fusion proteins and Geneious
Making MIT iGEM cookies