Lamitina Lab
at the University of Pittsburgh

Lamitina Lab Protocols

 

 

C. elegans methods

• CRISPR engineering of C. elegans

• G4C2 in situ hybridization

• Making M9

• Preparation of NGM agar plates

• Bleaching worms

• Freezing worms

• Lifespan assay

• Mutagenesis screen

• RNA isolation

• Genomic DNA isolation

• UV transgene integration

• Single worm PCR

• Microscopy on anesthetized worms

• C. elegans cell culture and FACS sorting

• Dye staining neurons

• PCR confirmation of single copy insertions (MOSci)

• Sytox staining C. elegans (live/dead)

• Isolation of organic osmolytes from C. elegans

• Making potassium phosphate solution for NGM plates

• Growing bacteria for C. elegans feeding RNAi

 

Molecular Biology Methods

• Rapid plasmid screening method ('Toothpick' method)

• dsRNA isolation from bacteria

• Bacterial transformation

• BCA protein assay

• Mammalian chromatin immunoprecipitation (CHIP)

• Ethanol precipitation of DNA

• Fosmid DNA isolation

• Freezing bacterial strains

• Glycerol assay

• Making PCR fusion products

• Phenol/chloroform DNA extraction

• qPCR setup

• qPCR worksheet template