Gene Pulser Electroporation Bu

Purification of Gene Targeting Vector DNA for Electroporation

and allow to it air dry. Resuspend the DNA in sterile TE (10 mM Tris-HCl, pH 8.0, 1.0 mM EDTA) at 2 mg/ml and deliver it to the Transgenic Core for electroporation. Prior to electroporation, we will verify the concentration and run it on a minigel to check

Stable Transfection (Electroporation)

medium. Supplies & Equipment: BioRad gene pulser with capacitance extender Electroporation cuvettes with 0.4 cm interelectrode distance  Reagents: sterile 1 x HeBS buffer (Hepes buffered saline) G 418 (Geneticin

Preparation of Microinjection Buffer

Buffer composition is 10 mM Tris-HCl, pH 7.5, 0.1 mM EDTA, 30 microM spermine, 70 microM spermidine, 100 mM NaCl. This buffer is more likely to produce transgenic mice with intact, unfragmented DNA molecules. See the following references: Schedl