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A Simple and Easy Method for RNA Extraction from the Cyanobacterium Synechocystis sp. PCC 6803
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DOI：10.21769/BioProtoc.5654.png)
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Abstract
En 中文
Cyanobacteria have been widely used as model organisms in photobiochemical research and have recently been exploited as hosts in numerous pilot studies to produce valuable biochemicals via genetic and metabolic modifications. Analyzing cellular RNA is a suitable method for studying genetic changes in cells. Several methods have previously been reported for cyanobacterial RNA extraction. However, the majority of these methods rely heavily on phenol and chloroform, which are hazardous. Additionally, these methods are time-consuming and difficult to perform. Using Synechocystis sp. PCC 6803 as a model, this study developed a novel method for extracting total ribonucleic acid (RNA) using standard centrifugation techniques and laboratory chemicals such as citric acid, ethylenediaminetetraacetic acid, sodium dodecyl sulfate, sodium chloride, and tri-sodium citrate dihydrate to extract RNA from cyanobacterial cells. This method does not necessitate the use of hazardous chemicals, especially phenol and chloroform. Furthermore, it is cost-effective since it does not require expensive chemicals. The results of the quantification, purity, and integrity checks show the effectiveness of this method for extracting good-quality RNA. Furthermore, RT-qPCR results demonstrate that the quality of the extracted RNA is suitable for downstream applications.
Keywords:
Cyanobacteria
Synechocystis sp. PCC 6803
RNA extraction
Quantitative reverse transcription polymerase chain reaction (RT-qPCR)
Single-stranded complementary DNA (cDNA)
Journal
B
IF:
1.1
Papers:
191
Citations:
5.4K
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Cited Papers