This page lists some standard protocols and recipies used by the Nebenführ lab. Some are very specific to our setup here in Knoxville, others are more generic. We hope that they are also useful for other people. And as always, if you notice a mistake in a protocol, or know of a better way of doing things, please let us know! (Note that all protocols are in PDF format.)

Molecular Cloning

• Standard PCR protocol

• Transformation of E. coli cells (heat shock)

• Making competent Agrobacterium tumefaciens cells; alternative version

• Transformation of Arobacterium tumefaciens (electroporation)

• Overnight cultures

Transformation of Plant Cells

• Transient expression after particle bombardment

• Stable transformation of BY-2 cells

Cell Culture

• BY-2 suspension culture medium + maintenance

• BY-2 transformation

• Immunofluorescence labeling of BY-2 cells with formaldehyde fixation or with glutaraldehyde fixation

Arabidopsis

• Germination medium & selection

• Plant DNA extraction