The Northern Blot

The Northern blot allows identification of specific messenger RNA sequences within a mixture of RNA molecules. The final signal achieved on the blot is proportional to the number of specific sequences present, allowing for a quantitative analysis of gene expression. The RNA transcripts produced from a particular gene can vary in size because of phenomena such as the following:

1. Utilization of a secondary transcription start site by RNA polymerase;

2. Premature termination of transcription resulting from nonsense mutation;

3. Posttranscriptional modifications, such as splicing;

4. Deletions within the gene coding sequence.

These alternative transcripts can be detected by Northern blot analysis. This information is routinely used to determine if the expression of a specific gene is altered in any way. Aberrations in gene expression are frequently studied in the laboratory to evaluate the cellular response to a particular stimulus or treatment at the molecular biology level. Because abnormal expression of specific genes is often reliably associated with certain disease states, the Northern blot can also be a valuable tool for diagnosis/prognosis in the clinical setting.

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