Project description:To identify genes affected by Pha13, Pha13 was transiently overexpressed in leaves of P. aphrodite by agroinfiltration. The leaves of P. aphrodite infiltrated with agrobacterium carrying empty vector (without Pha13) was used as a control. Transcriptional profiling was analyzed using a customized oligonucleotide array with RNA extracted from Pha13 and empty vector overexpressed leaves of P. aphrodite. The sequences used for probe design were collected from the Orchidstra 2.0 database (http://orchidstra2.abrc.sinica.edu.tw/orchidstra2/index.php). Totally, 42,973 probes were designed on this oligonucleotide array.
Project description:To identify genes affected by Pha21, Pha21 was transiently overexpressed in leaves of P. aphrodite by agroinfiltration. The leaves of P. aphrodite infiltrated with agrobacterium carrying empty vector (without Pha21) was used as a control. Transcriptional profiling was analyzed using a customized oligonucleotide array with RNA extracted from Pha21 and empty vector overexpressed leaves of P. aphrodite. The sequences used for probe design were collected from the Orchidstra 2.0 database (http://orchidstra2.abrc.sinica.edu.tw/orchidstra2/index.php). Totally, 42,973 probes were designed on this oligonucleotide array.
Project description:We performed one degradome sequencing for identification and characterization of novel microRNAs in Phalaenopsis aphrodite subsp. formosana. Plant tissues of leaves, stalk, and flower buds frozen in liquid nitrogen were ground to fine powder using a mortar and pestle, respectively. Total RNA extraction was freshly prepared using Trizol (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. About 50 μg of leaves with or without low temperature treatment, stalks and flower buds were taken separately and then mixed. The degradome library was constructed by the Genomics BioSci & Tech. Company following the manufactoring protocol. In brief, 200 μg of total RNA was passed though polyA column using the Oligotex kit (Qiagen). The 5'-RNA adapter containing a Mme I recognition site was ligated by T4 RNA ligase and further reverse transcribed to amplify the templates. After digesting with Mme I then ligated to a 3'-double DNA adapter, the products were amplified by additional PCR cycles and gel-purified for Illumina sequencing. The files contain the raw data of degradome sequencing. Examination of one degradome of mixed tissues of Phalaenopsis orchid
Project description:We performed one degradome sequencing for identification and characterization of novel microRNAs in Phalaenopsis aphrodite subsp. formosana. Plant tissues of leaves, stalk, and flower buds frozen in liquid nitrogen were ground to fine powder using a mortar and pestle, respectively. Total RNA extraction was freshly prepared using Trizol (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. About 50 μg of leaves with or without low temperature treatment, stalks and flower buds were taken separately and then mixed. The degradome library was constructed by the Genomics BioSci & Tech. Company following the manufactoring protocol. In brief, 200 μg of total RNA was passed though polyA column using the Oligotex kit (Qiagen). The 5'-RNA adapter containing a Mme I recognition site was ligated by T4 RNA ligase and further reverse transcribed to amplify the templates. After digesting with Mme I then ligated to a 3'-double DNA adapter, the products were amplified by additional PCR cycles and gel-purified for Illumina sequencing. The files contain the raw data of degradome sequencing.