Apple Virus Diagnosis Using Simplified RNA Extraction Method

사과바이러스 간편 진단을 위한 RNA추출법 개선

  • Shin, Dong-Il (Department of Biotechnology, Catholic University of Daegu) ;
  • Park, Hee-Sung (Department of Biotechnology, Catholic University of Daegu)
  • 신동일 (대구가톨릭대학교 생명공학과) ;
  • 박희성 (대구가톨릭대학교 생명공학과)
  • Received : 2009.10.08
  • Accepted : 2009.12.21
  • Published : 2009.12.31

Abstract

Kyungsan nursery complex which has a vast area for the production of various species of fruit tree stocks is in a high demand of virus-free saplings. Apple tree stocks, the most important products, urgently need more rapid and reliable viral diagnosis. In this study, a bead beater was tested because of convenience in dealing with large number of samples. Also, industrial glass bead abrasive (0.4 mm in diameter) at very low cost was used in a disposable way. For bead beater-aided RNA extraction from apple stem tissues, the guanidine thiocyanate method was confirmed to be very reliable. Silca membrane filter tube in connection to vacuum filtering device was strongly suggested for simplifying RNA capture and washing steps. Apple virus detection was confirmed by RT-PCR.

경산 묘목단지는 대단위 과수종묘를 생산하고 있어 경쟁력 강화를 위하여 무독묘 보증 생산이 요구되고 있다. 특히 생산규모가 큰 사과종묘에 대한 빠르고 정확한 바이러스 진단이 시급하다. 본 연구에서는 사과바이러스 진단을 위하여 다량의 시료를 동시 분쇄할 수 있는 bead beater를 이용하였으며 분쇄 bead는 저가의 산업용 glass bead (0.4 mm 직경)를 일회용으로 채택하였다. RNA추출을 위하여서는 guanidine thiocyanate 용액이 Trizol 용액보다 효과적인 것으로 나타났으며 silica membrane tube의 이용으로 RNA추출 간편성을 높일 수 있었다. 사과바이러스는 RT-PCR에 의하여 검증하였다.

Keywords

References

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