Note: These protocols are intended for lab members and are not meant to be complete stand-alone instructions.

Preparation and Transformation of Highly Efficient Chemically Competent E. coli

Soderling Lab Recombineering Protocol
(Copied from Liu, P., et.al., Genome Research, 13:476-484; 2003 with some changes.)

Site-Directed Mutagenesis
(Adapted from a combination of Stratagene’s QuikChange Site-Directed Mutagenesis Kit, and Wang, W.;Malcolm, B.A. 1999. Biotechniques. 26:680-682.)
        Site-Directed Mutagenesis (SupportProtocol1)
        Site-Directed Mutagenesis (SupportProtocol2)

Overlap Extension PCR
(1997. Biotechniques. 22:28-30.)

Calcium Phosphate Transfection

Lipofectamine Transfection of Neurons

Electroporation of Neurons

Expression and Purification of GST-Fusion Proteins
(Procedure Version 1: With Lysozyme)

Expression and Purification of GST-Fusion Proteins
(Procedure Version 2)

Purification of Tap-Tagged Proteins
This procedure has been optimized for the purification of AKAP-KL, but can be adapted and optimized for other proteins.

Silver Staining of Proteins in Polyacrylamide Gels

Fix - Actin Stain Protocol

Affinity Purification of Antibody From Antiserum

Reagents

Lab Recipes

IP Lysis Buffer