Protocols that are usually based on silica filters are not suitable for small RNA isolation. That is why several extraction and isolation kits still use both silica and phenol when aiming for isolating total RNA. As a result, the kits that do not contain phenol are changing their extraction chemistries, their filter, or both.
Some kits that come with silicon carbide resin will be a better option for isolating total RNA as well as small RNA. The major benefit of using silicon carbide is that it is capable of offering greater compassion for small RNAs. Another benefit is that it makes the process simpler. This means that you do not need to use phenol for your small RNA isolation process. This is for the reason that silicon carbide is capable of retaining the small RNAs much better than silica achieves.
Moreover, silicon carbide binds RNA at an equal rate, regardless of size. Additionally, when the RNAs are small, it binds them without partiality with respect to their GC content. Thus, there is no bias of improved RNAs. It has been revealed that for small RNAs, the grouping of phenol and precipitation-based separation can show discriminating loss of low-guanine-cytosine content miRNA, whereas no important prejudice has been observed for isolation methods using silica and phenol.
There are also miRNA Tissue Kits out there for small RNA isolation that use an automatic RNA-extraction system. These kits not only provide a look at the small RNAs but also at the complete range of RNA in a sample. They offer better coverage of the miRNA in the sample, which allows the users to regularize small RNA to the bigger messenger RNA.
Besides offering higher throughput, the automatic RNA-extraction system is particularly useful for researchers who are expecting for up-regulation or down-regulation of small RNAs. This is for the reason that the system offers them a depiction of all RNA species.
All kits that are used in total RNA extraction from tissues for analyzing the micro Ribonucleic acid and target gene expression are created equal. Micro Ribonucleic acids are short non-coding Ribonucleic acids that adjust gene expression. The abnormal expression of these nucleic acids is related to various diseases and they have both remedial as well as biomarker potential. However, the understanding of the usefulness of the microRNAs mainly counts on the tools users have to study them.
Earlier studies have recognized the need to optimize and standardize the methods of RNA extractionwith the intention of avoiding biased results. Researchers use five total RNA extraction methods that are commercially available to extracted RNA from murine lung, brain, and liver tissues. One of these methods of extracting RNA from tissues includes the use of phenol. It is either used to extract chloroform followed by alcohol precipitation or followed by solid-phase extraction.
Another method used for total RNA extraction is the solid-phase separation method with or without affinity resin. Then, each extraction is evaluated for the quantity and quality of RNA recovered, as well as the expression of target genes and miRNAs
It was finally concluded that chloroform phase isolation combined with the solid extraction method that is based on silica column was preferable.