Effectiveness of fluorescent dye DNA staining in simple DNA extraction experiments.

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  • 簡易ＤＮＡ抽出実験における蛍光試薬を用いたＤＮＡの検出とその有効性
  • カンイ DNA チュウシュツ ジッケン ニ オケル ケイコウ シヤク オ モチイタ DNA ノ ケンシュツ ト ソノ ユウコウセイ

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Abstract

<p>DNA extraction experiments have been carried out in many high school Biology classes. Although white extract thought to be DNA is obtained by the simple extraction methods described in textbooks, there are no validation experiments as to whether the white extract is DNA. The extracts from broccoli show the maximum wavelength of 260nm-specific of DNA, but the extracts from bananas did not. Bananas are reportedly not an appropriate material for this reason. The sensitivity of diphenylamine method specified with DNA detection was low, so we could not clearly confirm the existence of DNA in these extracts obtained by the simple extraction method. Therefore, we conducted a study to teach the experiments of DNA extraction and DNA detection within a 50-minute lesson period. And to detect the DNA from the sample, we used the fluorescent staining procedure with a fluorescent dye (GRG) specified for DNA detection. It was possible to observe strong fluorescence inthe extraction of broccoli, but little fluorescence in that of bananas. We could detect a clear difference in these two extracts. Then we introduced this fluorescent staining method in high school Basic Biology classes. Students could do all the experiments within 50 minutes. Most of the students were able to appropriately determine which extracts included DNA through the fluorescent staining procedure. These results suggest that fluorescent staining is an effective method for high school students to use to detect DNA from crude samples.</p>

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