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.
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.
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.