Manuals and Protocols

Here are the most recent manuals and protocols to clone and express shRNAs/miRNAs and cDNAs using the viral system. The protocols and manuals have a version number in order to keep track of the latest version. Each manual include detailed protocols to make lenitviruses and the maps of each vector. For the RNAi studies, the manual also include a detailed protocol on how to clone shRNAs/miRNAs.

Manuals

Manual for RNA interference studies: Version 2.2, November 2010 (old v.2.1 here).

Manual for Expression studies: Version 1.1, November 2010 (old v1.0 here).

 

Protocols

Transfection of 293T cells to produce lentiviruses (Version 3.1, October 2009).

Transfection of 293GAG/POL to produce retroviruses (Version 3.6, October 2009).

Crystal violet staining of cells (Version 2, August 2009).

General plasmid subcloning (Version 3.5, August 2009).

Partial digestions for plasmid subcloning (Version 1.0, November 2009).

General Geneclean protocol (Version 1.5, August 2009).

General transformation of E.coli (Version 2.0, August 2009).

Insertion of oligonucleotide adapters into plasmids (Version 1.5, August 2009).

Generation of whole cell extracts for SDS-PAGE (Version 1.0, October 2009).


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