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Composing solutions for spin-column based RNA purification from eukaryotic cells: An educational research article

Mahmoud M. Abdelfattah, Mohamed A. Elnagar, Mohamed F. Ibrahim, Magdy Albert, Maya M. Talal, Ehab El-Dabaa, Reham Helwa.


Twenty years ago, working with molecular biology research was very costly worldwide. Later on, after distributing the know-how of many reagents, it becomes more feasible to work in this field with reasonable costs. In addition, the optimization of large-scale production of enzymes, kits, lab consumables, and reagents made a big revolution in research and diagnostics. So, having affordable technologies in any molecular biology laboratory is an aim itself, especially in developing countries. After Covid-19 crisis, the need of molecular biology reagents is in surge. However, consumables’ prices, delayed orders, and shortage are the prominent issues after the pandemic. Thus, in the present study, homemade solutions were investigated for RNA purification from HepG2 and huh7 cells. The RNA was successfully purified by the homemade solutions and amplified using qRT-PCR. However DNA contamination was encountered which could be eliminated simply by DNase I digestion or designing proper primers in exon-exon junction for RT-PCR, in addition to precise normalization strategy.

Key words: RNA purification; silica-based columns; eukaryotes; guanidine thiocyanate; HepG2 cells.

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