Purification of RNA

KIT BASED PURIFICATION

Liquid- liquid purification often lead in contamination of RNA that's why researchers are switching towards solid based methods like silica fiber, glass beads methods and these approaches now available commercially in form of kits. They are not only easy to use but also more efficient and time saving.

For extraction and purification different kits are also available divide on the basis of RNA sample origin like from bacteria, yeast, cell line, mammalian cell and tissues so on and so forth.

Following both kit methods are used by Rodr?guez ML et al. in 2018.

DIRECT-ZOL RNA MINIPREP KIT:

This kit is based on tri-reagent system, this classical method for purification of RNA available in kit form but this kit is easier and efficient and where classical method achieve task in 90 minutes it only takes 7 minutes in complete purification.

Samples first homogenize with tri-reagents (guandine isothiocynate and phenol) then precipitated sample was treated with DNA I, rinse with RNase free water and passed through zymo-spin column which bind and elute RNA, no post purification method required for molecular analysis.

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RNeasy MINI KIT BY QUAIGEN:

This kit is specifically yeast RNA purification based on silica column. In this yeast cells are first converted into spheroplasts by digestion of cell wall of yeast with help of zymolase separated by centrifugation in salted buffer and ethanol for selective binding with membrane then directly loaded on RNeasy silica spin column. In end RNA remain bind with membrane while other biological contaminant elute out during washing.

Longer than 200 nucleotide yeast RNA molecules can be purified. Selective RNA elution can also made even without phenol presence.

Both kits has its own advantages and disadvantages. If we compare both kit protocols; RNA integrity provides by Rnesy min kit is better, having low toxicity due to absence of phenol but it is expensive and we cannot used them for filamentous fungus while direct-zol kit product is toxic quality of RNA is low but can run small samples provides high yield, low cost with no RNA loss.

CONCLUSION:

There are numerous protocols for purification of RNA, improvement in technology is a continuous process. But no single compound completely purify nucleic acid, multiple techniques are require for higher effectiveness in yield. But now silica column kit purification is mostly preferred by scientist as solid based method it can purify specific type of RNA, liquid-liquid purification method often result in molecular contamination while the appropriate selection of method should be based on the target and the system priorities.