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Status |
Public on May 05, 2021 |
Title |
StAgo1-GFP H2O 1 |
Sample type |
SRA |
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Source name |
Leaves
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Organism |
Solanum tuberosum |
Characteristics |
potato cultivar: StAgo1a-GFP (Sarpo Mira) dpi: 5 tissue: Leaves ip antibody: GFP (Chromotek, gtm-100)
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Treatment protocol |
H2O inoculations were added to the leaves 4 weeks post transfer to soil from in-vitro conditions.
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Growth protocol |
Potato plants were grown in-vitro at 22°C with a 16-h photoperiod on Murashige-Skoog (MS) medium (Duchefa Biochemie B.V., Amsterdam, Netherlands), supplemented with 2% (w/v) sucrose and 0.3% (w/v) gelrite (Duchefa). After 3 weeks plants were transferred to pots with soil in a growth chamber with the same light and temperature regime. P. infestans were maintained at 20 °C on rye agar medium supplemented with 2 % sucrose (Caten & Jinks 1968), rifampicin (10 μg ml−1), and pimaricin (10 μg ml−1). Transgenic lines of P. infestans isolate 88069 were maintained as above, but with the addition of the antibiotic geneticin (G418; 10 μg ml−1) to the medium.
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Extracted molecule |
total RNA |
Extraction protocol |
Inoculated leaves were harvested and flash frozen in liquid nitrogen. StAGO1a-GFP and control Pi-GFP immunoprecipitations were performed with anti-GFP antibody (Chromotek, gtm-100). Barcoded sequencing libraries were constructed using the Ion Total RNA-Seq Kit v2 and Ion Xpress barcode primers. Sequencing was performed on the Ion Proton platform using the Ion PI™ Hi-Q™ Sequencing 200 Kit (Thermo Fisher Scientific).
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Library strategy |
miRNA-Seq |
Library source |
transcriptomic |
Library selection |
size fractionation |
Instrument model |
Ion Torrent Proton |
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Data processing |
smallRNA processing: The function SamToFastq, included in the Picard tool package, was used to convert the original BAM format to FASTQ format (for the raw data which was not already in FASTQ format), on which a quality control was performed, using FastQC smallRNA processing: Cutadapt, quality trimming (20), read length filtration 18 - 38 nt smallRNA processing: Bowtie2 in combination with SAMtools view were used for tRNA and rRNA trimming and potato and P. infestans sRNA filtering to exclude the reads from the other genome for the different datasets, using 0 mismatches allowed per seed against each Fasta filter smallRNA processing: SamToFastq converted the files to fastq smallRNA processing: Seqtk was applied to convert the FASTQ files to FASTA. Degradome processing: Raw RNA reads from degradome libraries were adaptor- and quality trimmed with Cutadapt Degradome processing: Seqtk was applied to convert the FASTQ files to FASTA. The sRNA and degradome fasta files and were analyzed in PAREsnip2. PAREsnip output data was imported in R where PAREsnip2 replicates were merged and duplicated targets were discarded. Resulting datasets were analyzed in our machine learning script (https://github.com/kristianHoden/degradome). False positives were discarded The datsets were sorted in list according to their Normalized Fragment Abundance (NFA) and targeting transcriptome Solanom tuberosum genome v4.04, Phytophthora infestans genome (ASM14294v1) fasta format of small RNA and degradome libraries Genome_build: Solanom tuberosum genome v4.04, Phytophthora infestans genome (ASM14294v1) Supplementary_files_format_and_content: pinfDown - contain the datasets with targets in P. infestans with decreased NFA of the targets upon infection Supplementary_files_format_and_content: pinfUp - - contain the datasets with targets in P. infestans with increased NFA of the targets upon infection Supplementary_files_format_and_content: potDown - contain the datasets with targets in potato with decreased NFA of the targets upon infection Supplementary_files_format_and_content: potUp - - contain the datasets with targets in potato with increased NFA of the targets upon infection Supplementary_files_format_and_content: rDown - contain the datasets with targets in resistance (R) genes in potato with decreased NFA of the targets upon infection Supplementary_files_format_and_content: rUp - contain the datasets with targets in resistance (R) genes in potato with increased NFA of the targets upon infection
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Submission date |
Dec 16, 2020 |
Last update date |
May 05, 2021 |
Contact name |
Kristian Erik Persson Hodén |
E-mail(s) |
kristian.hoden@gmail.com
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Organization name |
Swedish University of Agricultural Sciences
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Department |
Plant Biology
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Lab |
Christina Dixelius
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Street address |
Almas allé 5
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City |
Uppsala |
ZIP/Postal code |
75651 |
Country |
Sweden |
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Platform ID |
GPL29506 |
Series (1) |
GSE163382 |
Convolutional neural network modelling: advancing identification of true mRNA cleavage sites |
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Relations |
BioSample |
SAMN17098845 |
SRA |
SRX9694812 |
Supplementary data files not provided |
SRA Run Selector |
Raw data are available in SRA |
Processed data are available on Series record |
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