title: Plant virus detection microarray
status: Public on Jun 30 2010
contact.city: Beijing
contact.first: Yongjiang
contact.last: Zhang
contact.email: zhangyjpvi@yeah.net
contact.country: China
contact.institute: Chinese Academy of Inspection and Quarantine
organism: Sobemovirus
  Carlavirus
  Capillovirus
  Potexvirus
  Potyvirus
  Tobamovirus
  Comovirus
  Nepovirus
  Tobravirus
  Cucumovirus
  Ilarvirus
  Pomovirus
  Orthotospovirus
description: 70mer probes were designed to detect plant viruses infection in genus level. This microarray platform is able to detect 169 plant virus species of 13 virus genera.
manufacture_protocol: The 70-mer oligonucleotide arrays were synthesized on 75-mmx25-mm glass slides using a SmartArrayer system (CapitalBio, China). All probes were spotted three times on the microarray. The reverse complementary sequences of the Capillovirus and Tospovirus probes were synthesized to analyze the probe strand effect. Three types of control probes were spotted. The 5¡¯ amino-linker oligonucleotide (5¡¯- TTTTTTTTTTTTTTTCTCATGCCCATGCCGATGC -3¡¯) was used as the positive control. This positive control was a random sequence which did not hybridize with any virus sequences. It was mixed with the samples before hybridization, and used to monitor the hybridization of the microarray. The spotting buffer was used as the negative control. The internal control was 5¡¯ Hex-linker oligonucleotide (5¡¯- GTCACATGCGATGGATCGAGCTCCTTTATCATCGTTCCCACCTTAATGCA -3¡¯). The internal control was spotted on the microarray to monitor the ligation of probes to the chips. Spotted slides were air dried for 10 min and then fixed by UV exposure at 240mJ/cm2. After being incubated in a 0.5% sodium dodecyl sulfate solution at 60¡æ for an hour, slides were rinsed in 100 mM Tris (pH 8.0) for 5 min, and kept moist at 4¡æ for use.