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Analysis of microRNAs by Northern Blot Technology (CAT#: STEM-MHT-0130-LGZ)

Introduction

MicroRNAs (miRNAs), non-coding regulatory RNA molecules approximately 21 nucleotides in length, represent a new dimension of post-transcriptional regulation of gene expression. The in-depth study of miRNA requires the development of effective miRNA detection methods, such as northern analysis, quantitative real-time PCR, and microarray. Northern analysis is a widely used method of miRNA analysis because it is usually a readily available technique in laboratories and does not require special equipment and technical knowledge.




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.