Lamitina Lab
Protocols
Worm Methods
- Worm total RNA isolation (Qiagen/RNAeasy method)
- ENU mutagenesis (F2 screen)
- UV mediated transgene integration
- Live/dead staining with Sytox
- Osmolyte isolation from worms
- Lifespan Assay
- Anesthetized worm microscopy
- Cell culture and FACS sorting
- Single Worm PCR
- Worm genomic DNA prep
- OSM worksheet
- OSR worksheet
- PCR confirmation of single-copy insertions
RNAi screening
- RNAi library 384-96 well expansion worksheet
- RNAi colony streaking worksheet
- Bacterial cultures and plate seeding worksheet
- Plates Screened Worksheet
- Screen log sheet
Solutions and recipes
- Potassium phosphate buffer (for NGM plates)
- NGM plate recipe - isotonic and hypertonic NaCl
- 5X Gel loading buffer and DNA ladder recipes
Molecular Biology
- Bacteria transformation
- DNA precipitation
- Phenol:Chloroform DNA extraction
- BCA protein assays (for glycerol assay normalization)
- One-tube BP/LR Gateway reaction
- Growing RNAi bacteria
- qPCR assay setup
- qPCR plate template
- PCR fusion
- DNA sequencing form
- Gateway PCR primer design
- Gateway BP reaction setup
- Bacterial cracking for rapid plasmid analysis