What is Freund's adjuvant and Tissue RNA isolation?

Freund's adjuvant is used as a booster and it is a solution, in which antigen is emulsified in mineral oil. It is of two types namely FCA or Freund's Complete Adjuvant or FIA or Freund's Incomplete Adjuvant. While the complete form is composed of inactivated and dried mycobacterial components, generally, M. tuberculosis, the incomplete form does not contain inactivated and dried mycobacteria. It is named after an immunologist, who was born in Hungary and raised in America, Jules T. Freund.

The complete adjuvant is effective in motivating cell-mediated immunity and it is capable of increasing the powerfulness of T helper cells in the manufacture of immunoglobulins as well as effector T cells. Regulatory authorities prohibit its use on the human body, owing to its toxicity. Currently, there are guidelines related to its use even for animal research, owing to its hurting reaction and possibility for the damage of tissues.

Injections of Freund's Complete Adjuvant should be subcutaneous, as intradermal injections may result in ulceration and necrosis of the skin. Temporary or permanent muscle lesion may be caused due to intramuscular injections. Intravenous injections may cause pulmonary lipid embolism.

When considering the optimistic effects of Freund's adjuvant, it is found that its complete form has the ability to prevent juvenile-onset diabetes in non-obese diabetic mice. It is capable of reversing diabetes when it is used by combining it with spleen cells. It is also established that even without combining FCA with spleen cells, it has the ability to restore insulin-manufacturing beta cells in the pancreas of these mice. However, the reverse of end-stage diabetes is possible by combining spleen cells with FCA.

Tissue RNA isolation is an essential forerunner to different molecular and genomic biology applications, such as in next-generation screening, sequencing, and gene expression analyses. The structural intricacy of eukaryotic tissues and cells, as well as the feeling of RNA molecules, may pose technical hurdles during the process of extraction. However, several ready-to-use commercial kits and reagents are now available. They are specifically designed for purifying the RNA tissue through the vacuum, spin, or magnetic-based techniques. Even some kits have automation compatibility, as well.

Tissue RNA isolation has been tested on animals having Parkinson's disease while analyzing their gene expression. Efficient interference and homogenization of animal tissues are necessary to guarantee a high yield of RNA. While interference releases RNA, homogenization decreases sample viscosity to make RNA purification easy.

Many extraction kits are available with tools that use high-tech ultrasound technology to disrupt and homogenize tissues efficiently in a single step. Each of these RNA extraction kits comes with an RNA extraction reagent and it is used as a sonication medium. It maintains the reliability of RNA whilst disrupting cells and dissolving cell constituents. Some of the unique benefits of using the RNA extraction reagent with other agents for tissue disruption and homogenization include:

·         Fast protocol

·         Non-contact reduces pollution

·         Isothermal process

·         Resourceful and reproducible

·        Multiplexing ability of more than one sample in parallel

When different methods are used for Tissue RNA isolation, it will yield diverse results and thus, a healthy RNA isolation technique is necessary for reproducibility.