Abstract
We use the Miller chromatin spreading technique for electron microscopic visualization of transcriptionally active genes. In this method, cells are hypotonically disrupted and cellular contents are diluted into water at pH 8-9 and fixed with formaldehyde. The dispersed cellular contents are centrifuged onto a carbon-coated EM grid; the majority of the material that is deposited on the grid consists of entangled masses of dispersed chromatin, some regions of which are transcriptionally active (Fig 1). Our interests lie in ultrastructural analysis of co-transcriptional RNA processing events on pre-mRNA transcripts, which we analyze by mapping structural features on successive nascent transcripts on a given gene. The two processing events that we have been able to study by this approach are the removal of introns by splicing and generation of the 3’ end of the transcript.
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