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Analysis of Plant Mitochondria RNAs by Northern Blot Technology (CAT#: STEM-MHT-0133-LGZ)

Introduction

Transcriptome analyses enhance our understanding of RNA­based regulatory mechanisms in prokaryotic organisms as well within the nuclear­cytosolic compartments of eukaryotic cells. Higher plant organelles employ complex post­transcriptional regulatory mechanisms, which are studied in a comprehensive fashion by different RNAmethodologies. Northern blot hybridization provides with important data to the "steady­state" level of accumulation of a given transcript in the sample, and thus remains as a standard method for detection and quantitation of RNA levels, despite the advent of other powerful techniques.




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

Gene 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.