Protocols

Protocols

Gene Expression Methods

Nuclei isolation and nuclear run-on assay

Broccoli (Brassica oleracea) RNA polymerase I holoenzyme purification

Promoter-independent RNA polymerase activity assay

Promoter-dependent in vitro transcription and S1 nuclease protection assay (for Pol I)

 

Chromatin and DNA Methods

Purification of core histones from Broccoli (Brssica oleracea)

 

Cytogenetics and Cell Biology Methods

Chromosome counting in DAPI-stained anther filaments

 

 Antibody methods

Affinity purification of anti-peptide antibodies 

 

Agrobacterium tumefaciens methods

Agrobacterium growth and transformation by Freeze/thaw shock or electroporation to introduce plasmid DNA vectors

Agrobacterium tumefaciens plasmid DNA mini-prep

 

About the banner image (top of page)

The dark color of soybeans (middle) is extinguished in cultivated varieties (left) because of natural posttranscriptional silencing of the CHS gene and can be partially reversed by infection of the parental plant with a virus possessing a PTGS suppressor protein, producing a mottled pattern (right). Pikaard CS, O Mittelsten-Scheid (2014). Epigenetic regulation in plants. In: Epigenetics; edited by D Allis, T Jenuwein, D Reinberg. Cold Spring Harb Perspect Biol. 6(12):a019315. doi: 10.1101/cshperspect.a019315.