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Analysis of Small RNA species in Drosophila Melanogaster by Northern Blot Technology (CAT#: STEM-MHT-0137-LGZ)

Introduction

The last decades have witnessed the explosion of scientific interest around gene expression control mechanisms at the RNA level. This branch of molecular biology has been greatly fueled by the discovery of noncoding RNAs as major players in post-transcriptional regulation. Such a revolutionary perspective has been accompanied and triggered by the development of powerful technologies for profiling short RNAs expression, both at the high-throughput level (genome-wide identification) or as single-candidate analysis (steady state accumulation of specific species). Although several state-of-art strategies are currently available for dosing or visualizing such fleeing molecules, Northern Blot assay remains the eligible approach in molecular biology for immediate and accurate evaluation of RNA expression.




Principle

The RNA separated by agarose gel electrophoresis is transferred to a solid-phase carrier by a certain method, and then hybridized with a labeled probe, and the transcriptional expression can be quantitatively or qualitatively analyzed through the hybridization result.

Applications

RNA Analysis

Procedure

1. RNA extraction and quality control.
2. Denaturing gel RNA electrophoresis.
3. Transfer the membrane.
4. Probe synthesis and DIG labeling.
5. Prehybridization.
6. Hybridization.
7. Wash the membrane.
8. Data processing.

Materials

Samples of plants and animals provided by customers must be shipped on dry ice. If the RNA sample is provided, the following conditions must be met: the RNA integrity is good (28S, 18S bands are clearly visible), no degradation or DNA contamination, and the customer needs to provide the electrophoretic map; High RNA purity (OD260/OD280=1.9~2.1), customers need to provide spectrophotometric detection data; The RNA should be dissolved in double steaming water, the RNA concentration should not be less than 0.8 μg/μl, it is best to provide the amount of 2 experiments; Long distances need to be transported with dry ice, or RNA contained in ethanol with regular ice packs.