LOSS OF AN ABUNDANT QUANTITY OF RIBONUCLEIC ACID DURING MINI COLUMN ISOLATION METHOD

Abstract

Aim: The Isolation of nucleic acid is an important step for conducting different molecular assays in many laboratories around the world. It is also a common practice that user is isolating the ribonucleic acid (RNA) from the samples with mini column once and throwing away the supernatant. This makes isolated RNA as limiting factor in many studies as this issue has not been addressed in literature. Therefore, we decided to conduct whether it is a loss of ribonucleic acid during the mini column isolation method. Method: In this research, the mini column isolations were done with different samples of human tissues from placenta and umbilical cords and subsequent isolations of supernatants. Yields and successful isolations of RNA were assessed with spectrometric instrument and real time PCR machine. Results: It was found that there is loss of abundant quantity of RNA during the subsequent isolations. The amount measured with UV spectrometer indicates that some times 2nd and 3rd isolation have more RNA than the first isolation. Realtime PCR for house keeping gene beta actin shows that presence of RNA can be seen up to 6 isolation cycles from supernatants. Conclusion: There is loss of RNA in subsequent isolations with mini column method, therefore it is possible to isolate more RNA from subsequent supernatant isolations. User should do the multiple isolations to get higher yield of RNA.