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  [in Japanese] Effectiveness of fluorescent dye DNA staining in simple DNA extraction experiments.  [in Japanese]

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Abstract

<p>高等学校生物において,多くの高等学校でDNA抽出実験が実施されてきた.DNAを確認するためのジフェニルアミン法(馬場ら2013)は,感度が低いため,簡易抽出法で得られた抽出物では,はっきりとした結果が得られなかった.そこで, DNAの検出感度が高い確認方法で,簡便であり,授業1時間(50分)でDNAの抽出実験と確認実験の両方が実践できることを目的として研究を行った.今回,蛍光試薬(GRG)を用いた確認方法(蛍光試薬法)で,簡易DNA抽出実験で得られた抽出物の確認を行ったところ,バナナでは蛍光がほとんど見られず,ブロッコリーでは強い蛍光が見られ,これら二つの抽出物のDNA含有量の明瞭な違いを確認できた.この方法を用いて授業実践を行ったところ,1時間以内でDNAの抽出実験と確認実験を行うことができた.そして,多くの生徒は抽出物中のDNAの有無を適切に判断できるようになった.以上のことから,蛍光試薬法は生徒実験における簡便なDNA確認法として有効な方法であることが示唆された.</p>

<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>

Journal

  • Japanese Journal of Biological Education

    Japanese Journal of Biological Education 56(1), 21-28, 2015

    The Society of Biological Science Education of Japan

Codes

  • NII Article ID (NAID)
    130007719583
  • NII NACSIS-CAT ID (NCID)
    AN0012957X
  • Text Lang
    JPN
  • ISSN
    0287-119X
  • NDL Article ID
    026831842
  • NDL Call No.
    Z18-53
  • Data Source
    NDL  J-STAGE 
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