Total RNA Analysis and Isolation

RNA simply stands for ribonucleic acid and is present in all living cells. It carries instructions from deoxyribonucleic acid (DNA) to aid in protein synthesis, and in some cases, RNA also carries genetic information. It is a molecule with a sequence of chains. The bases of RNA are Adenine, Guanine, Uracil, and Cytosine. DNA goes through a process called transcription in which RNA is formed, and has different types. The difference occurs due to the varying encoding of the gene.

What is total RNA?

Total RNA, in simple terms, is all the ribonucleic molecules that occur in a specific cell. Basically, all the types of RNA fall under this category. There are four main types of RNA namely; Messenger RNA, Transfer RNA, Ribosomal RNA and Small Nuclear RNA. In deeper fields, it refers to ribonucleic acid that has been isolated from the body cells, to be used in genetic mapping.

How is total RNA obtained?

Through a process known as RNA isolation, total RNA is extracted from tissues and cells. RNA isolation is a keen and delicate process that can be done in various methods, following important steps. Cell disruption – this is the most vital step in the process and it can be done by either gentle or harsh methods. The most appropriate technique should be used to ensure that the best results are yielded. Scientifically, there are five major ways cell extraction can be done;

• Mechanical Homogenization
• Chemical Lysis
• Ultrasonic Homogenization
• Temperature treatments
• Pressure Homogenization

What is Total RNA Isolation?

This is a process that involves laboratory methods to obtain all the cell ribonucleic acid from a specific tissue. The technique extracts all ribonucleic types and not a specific RNA like other processes. It is important and beneficial for this process to take place. The extract is used in research and helps in getting better results and conclusions over time, as the protocol is repeated. Consequently, it helps in obtaining and studying better quality RNA.

Various chemical agents are used in the process to lower potential damage risks to the sample. A specific chemical should be chosen, however, to avoid unstable and degrading results. The techniques normally involve a stepwise protocol. First and foremost, a tissue sample is prepared, ensuring not to damage it. It is then treated using the chemical agents and the total RNA is finally isolated. One should do proper research and follow the rules strictly during the process.

Total RNA Sequencing

In some cases, there is need to analyze all the transcripts of a particular organism. This is whereby all the RNA types are researched on and explored. This is a great benefit as it not only covers the coding part of the genome but also the non-coding part of it. This process allows the researchers to capture all known and hidden features, observation at a wide range, identification of biomarkers in a really wide range of transcripts and the sample is well understood and explored.

Any process, method or technique involved in isolating, extracting or handling total ribonucleic acid should be done with caution and precision. There are manuals and kits that aid in making the processes more comprehensive.