Question

What are northern blots based on?

Answer 1

Northern blot:

1. It is a qualitative analysis to study ribonucleic acids (RNA) for gene expression analysis.
2. This blotting technique operates based on size-dependent RNA separation and helps identify the RNA transcript size.
3. It conducts RNA segregation based on size by gel electrophoresis and the separated RNA strands are identified using hybridization probes.
4. RNA is negatively charged and migrates towards the anode during gel electrophoresis.
5. The length of the RNA strand influences its migration speed where longer strands move slower compared to shorter strands.
6. This technique helps in identifying overexpression of oncogenes and determining the molecular weight of the RNA strands.
7. In northern blotting, the RNA of interest is extracted and isolated. This RNA molecule is then subjected to gel electrophoresis.
8. The RNA molecules are separated based on their size and they are transferred to a blotting membrane and immobilized on the membrane.
9. A labeled probe is added to the RNA mix and then visualized.