A Study on RNA Determination by Ribose Estimation -Condition of Perchloric acid Concentration on the Color generation by Orcinol reaction-

RNA 정량법(定量法)에 관한 연구(硏究) -과염소산(過鹽素酸)농도가 Orcinol 반응(反應)에 미치는 영향(影響)-

Effects of perchloric acid (PCA) and trichloroacetic acid (TCA) on the color generation by orcinol reaction were systematically investigated. When the concentration of PCA on hot acid treatment was varied from 1 through 5 to 10%, and then the concentration of PCA on orcinol reaction was adjusted to 5% of reaction volume, no difference in the color generation was observed between 5 and 10% of PCA, but clearly observed between 1 and 5% PCA. When RNA was treated in 5% hot PCA and then PCA concentrations on orcinol reaction were adjusted to 5% and 10%, respectively, remarkable differences in color generation were observed. When RNA was treated in 10% hot PCA and then PCA concentration on orcinol reaction was adjusted to 5% no difference in color generation between 5% and 10% hot acid treatment was observed. The results show that PCA concentration must be adjusted prior to orcinol reaction.

1) 열산처리(熱酸處理)에서 산(酸)의 농도(濃度)는 TCA, PCA 모두 5%이면 충분하다. 2)열산처리시간(熱酸處理時間)은 발색시간(發色時間)을 15분(分)으로 할 때 20이면 충분하다. 3) 20 분간(分間) 열산처리(熱酸處理)하면 발색시간(發色時間)은 15분(分)이면 된다. 4) PCA 농도(濃度)가 5% 이상(以上)에서는 RNA 전처리(前處理)에서는 영향을 미치지 않으나 발색(發色)시킬 때는 영향을 미치므로서 이때 PCA의 농도(濃度)를 일정(一定)하게 해줄 필요(必要)가 있다. 5) 발색(發色)시킬 때 시료(試料) 중(中)에 TCA가 5% 이상(以上) 공존(共存)하면 현탁되어 OD 측정(測定)이 불가능(不可能)하며 PCA처럼 농도차(濃度差)가 색도(色度)에 영향을 미칠 가능성(可能性)이 있으므로 열산처리(熱酸處理)는 TCA보다 PCA를 사용(使用)하는 것이 좋다.