|Table of Contents|

Optimization of Hybridization Condition for Lily Virus Detection Microarray(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2015年21
Page:
110-115
Research Field:
Publishing date:

Info

Title:
Optimization of Hybridization Condition for Lily Virus Detection Microarray
Author(s):
JIA Hui1; ZHENG Jie1; CAO Zhiyan1; WANG Jinzhong2; DONG Jingao1JIA Hui1; ZHENG Jie1; CAO Zhiyan1; WANG Jinzhong2; DONG Jingao1
（1.College of Life Science，Agricultural University of Hebei，Baoding,Hebei 071000; 2.Beijing University of Agriculture，Beijing 102206)
Keywords:
Lily virus; oligonucleotide microarray; hybridization conditions; optimization
PACS:
-
DOI:
10.11937/bfyy.201521029
Abstract:
In the present study the lily infected by Lily symptomless virus（LSV），Cucumber mosaic virus(CMV)，Lily mottle virus(LMoV) and tobacco infected by Tobacco mosaic virus (TMV)，Potato virus Y（PVY) were chosen as test material.Based on conserved coat protein region nucleotide sequences of five kinds of viruses，primers and probes were designed and the microarray were designed and fabricated.Total RNA of infected plant was extracted by the Trizol regent，and products that Cy3 labeled RT-PCR amplification hydrid with the microarray.The effects of whether the PCR products were denatured，hybridization time，hybridization temperature，hybridization buffer and the ratio of non-market primer to market primer with fluorescence in PCR system on microarray hybridization were evaluated.The result showed that the optimal hybridization time was 60 minutes；temperature was 42℃；SSC concentration was 6×SSC，SDS was 0.2%；The ratio of non-market primer to market primer with fluorescence was 1∶10 in the PCR system，and the PCR products must to be degenerated before hybridization.After the overall optimization，the gene chips had a high specificity in the hybridization，and could be used for detection of lily viruses fast and accurately.

References:

 

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Last Update: 2015-12-28