|
| Status |
Public on Nov 01, 2019 |
| Title |
5Pi_tZ_R1 |
| Sample type |
SRA |
| |
|
| Source name |
roots
|
| Organism |
Arabidopsis thaliana |
| Characteristics |
ecotype: Columbia
treatment: grown without phosphate and treated with trans-zeatin
|
| Growth protocol |
Arabidopsis thalinaan (Col-0) seedlings were grown in Murashige and Skoog medium containing 625 or 5 µM of phosphate for 8 days and then treated with 5 µM of cis-zeatin or trans-zeatin fro 8 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from Arabidopsis plants using the Trizol reagent and RNeasy columns (Qiagen). Quantity and quality measurements were taken using a NanoDrop spectrophotometer and Bioanalyzer.
After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. After agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used in quantification and qualification of the sample library. At last, the library could be sequenced using Illumina HiSeqTM 2000 or other sequencer when necessary
polyA library
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina HiSeq 2000 |
| |
|
| Description |
grown without phosphate and treated witht trans-zeatin
|
| Data processing |
Basecalls performed using CASAVA version 1.4
Clean (FastQC software (Babraham-Bioinformatics, 2014). Reads are mapped to a reference sequences using HISAT2 with 8 mismatches allowed.
Reads count to a reference genome (TAIR 42.gff3) using the htseq-count tool.
Differential expression analysis was calculated with edgeR with the following parameters: FDR<0.05, fold change >2 fold or < -2 fold
Genome_build: Atabidopsis thaliannTAIR 10
Supplementary_files_format_and_content: tab-delimited text files including normalized counts.
|
| |
|
| Submission date |
May 11, 2019 |
| Last update date |
Nov 01, 2019 |
| Contact name |
Juan Carlos del Pozo |
| E-mail(s) |
pozo@inia.es
|
| Organization name |
Centro de Biotecnología y Genómica de Plantas
|
| Department |
Biotechnology
|
| Lab |
180
|
| Street address |
Campus de Montegancedo, UPM
|
| City |
Pozuelo de Alracon |
| State/province |
Madrid |
| ZIP/Postal code |
28223 |
| Country |
Spain |
| |
|
| Platform ID |
GPL13222 |
| Series (1) |
| GSE131056 |
Role of cis-zeatin in root responses to phosphate starvation |
|
| Relations |
| BioSample |
SAMN11625717 |
| SRA |
SRX5822286 |
