Pikaard Lab Protocols

Gene Expression Methods

• Nuclei isolation and nuclear run-on assay

• Broccoli (Brassica oleracea) RNA polymerase I holoenzyme purification (PDF file)

• Promoter-independent RNA polymerase activity assay (PDF file)

• Promoter-dependent in vitro transcription and S1 nuclease protection assay (PDF file)

• Transient expression in Arabidopsis protoplasts (PEG method)

Chromatin Methods

• Chromatin immunoprecipitation and ChIP-chop (PDF file)

• Bisulfite-mediated DNA sequencing for mapping methylated cytosine positions (PDF file)

• Pulsed-field gel electrophoresis of Arabidopsis chromosomal DNA (PDF file)

• Purification of core histones from Broccoli (Brssica oleracea) (PDF File)

Cytogenetics and Cell Biology Methods

• Chromosome counting in DAPI-stained anther filaments

• Olga Pontes' protein immunolocalization and DNA-FISH methods (PDF file)

Nucleic Acid Purification

• Isolation of genomic DNA and total RNA from same total nucleic acid prep

• Single leaf DNA prep for Southern blot analysis of transgenic plants in first generation

Antibody methods

• Affinity purification of anti-peptide antibodies (PDF file)

Agrobacterium tumefaciens methods

• Freeze/thaw shock or electroporation of plasmid DNA into Agrobacterium tumefaciens

• Agrobacterium tumefaciens plasmid DNA mini-prep