Disclaimer: None of the authors, Mei Lab, or Case Western Reserve University assume any liabilities for these protocols. Use at your own risk. Our goal for these protocols is to disseminate open source techniques commonly used in the Mei Lab. However, these protocols do not assume any guarantees of correctness and should be implemented using safety protocols, provisions and permissions of the universities with which the users are operating within. Lastly, these protocols are distributed in the hope that it will be helpful, however without any warranty or desire of merchantability.

It is requested that any use of these protocols be cited in methods sections as “Mei Lab Protocols” + “Specific Protocol Name” along with website “meilab.org/protocols”.

Biochemical

• Hot Shot DNA Extraction

• Recombinant AAV Virus Production

• Lentivirus Packaging

• BrdU Injection

• Competent Cells

• Subcellular Fraction Isolation

• Western Blot

• Molecular Cloning

• Generation of CRISPR-Cas9 Plasmid and KO cells

Electrophysiological

• Slice Cutting

• Intracellular Solution

• mEPP and CMAP Recording

In Vitro

• Primary Neuron Culture

• Primary Microglia Isolation

• Elisa protocol for Lrp4 Agrin

• In Situ Hybridization

• Neurosphere Culture

• Immunofluorescence Staining of Whole-mount Mouse Diaphragms

• Perfusion and Staining with Stocks

In Vivo

• Fast Scanning Cyclic Voltammetry

• Denervation Nerve

• In Utero Electroporation