Experimental Molecular Genetics

Labs

WARNING NOTICE

The experiments described in these materials are potentially hazardous and require a high level of safety training, special facilities and equipment, and supervision by appropriate individuals. You bear the sole responsibility, liability, and risk for the implementation of such safety procedures and measures. MIT shall have no responsibility, liability, or risk for the content or implementation of any of the material presented.

Background

C. Elegans Anatomy (PDF)

C. Elegans Life Cycle (PDF)

Protocols

Transferring Worms (PDF)

RNAi Feeding Protocol (PDF)

Worm Lysis for PCR (PDF)

SNP Mapping Protocol (PDF)

Agarose Gel Electrophoresis (PDF)

Synchronization of C. elegans (PDF)

Chemotaxis Assay (PDF)

RNA Extraction (PDF)

cDNA Synthesis (PDF)

Cleaning Contaminated Worm Stocks (PDF)

Molecular Cloning using the Gibson Assembly Method: See the descriptions and protocols in the Gibson Assembly Cloning Kit Manual (PDF - 1.3MB), and the Gibson Assembly cloning kit (NEB E5510S), from New England Biolabs.

General Guidelines for Primer Design (PDF)

Resuspending PCR Primers (PDF)

Making Bacterial Glycerol Stocks (PDF)

Real-time PCR: This protocol draws from the following sources: Fraga, D., Meulia, T., et al. Unit 10.3 Real-Time PCR. Current Protocols, Essential Laboratory Techniques. Wiley Online Library, 2014.

Real-Time qRT-PCR. National Center for Biotechnology Information.

Analyzing Real-time PCR Data by the Comparative CT Method: This protocol is described in Schmittgen, T. D., and K. J. Livak. “Analyzing Real-time PCR Data by the Comparative CT Method.” Nature Protocols 3, no. 6 (2008): 1101–8.