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| Status |
Public on Jun 15, 2021 |
| Title |
AX4_FD_r1_hr24 |
| Sample type |
SRA |
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| Source name |
AX4
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| Organism |
Dictyostelium discoideum |
| Characteristics |
strain: AX4
phenotype group: WT
replicate: 1
developmental time: hr24
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| Treatment protocol |
Cells were washed with PDF and plated on nitrocellulse filter on filter paper saturated in PDF at 1.8x106 cells/cm2.
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| Growth protocol |
Dictyostelium discoideum cells were grown in nutrient media (HL-5)
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| Extracted molecule |
polyA RNA |
| Extraction protocol |
At each time point, we harvested the cells directly into 1 mL of Trizol® (life technologies, CA, USA) and extracted total RNA according to the manufacturer’s recommended protocol. We performed two rounds of poly-A selection and fragmented 100 ng of the resulting mRNA into approximately 200 bases fragments. We prepared cDNA and the second strand.
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| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina Genome Analyzer II |
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| Description |
GSM1517239
GEO: GSE61914
Filter development, replicate 1, hour 24
AX4_FD_r1_hr24
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| Data processing |
After demultiplexing and filtering illumina adapter sequences, the data was mapped to the D. discoideum genome with bowtie version 1.0.0 through the web application dictyExpress GenBoard or Genialis platform. Read qualities were used. Other parameters: seed length = 28, allowed mismatches = 2, allowed up to 1 alignment per read, with bowtie options -a, --strata, --best. Unmapped reads were trimmed from 3' by 2 nucleotides, which was repeated 5 times.
Raw expression values were computed as the number of reads that were uniquely mapped to gene exons.
Normalized expression were scaled with mappable exon lengths. This normalization is similar to the RPKM normalization (Reads Per Kilobase of exon per Megabase of library size), but instead of dividing by exon lengths we used the uniquely mappable parts of the exon. To obtain uniquely mappable parts, all possible subsequences of the reference genome (of the same length as reads in raw data) are mapped back to the reference genome, obtaining the number of uniquely mapped sequences to the exons (Exon_mappable). As a library size we used the total number of all uniquely mapped reads from the experiment, excluding the non-polyadenylated genes (N_unique). Normalized expressions were computed as follows: Exp = 10^9*raw/(N_unique * Exon_mappable).
Genome_build: D. discoideum Chromosomal DNA: 1,2,3,4,5,6,M, and floating contigs (created: 05-13-2009 13:53) from the DictyBase (chromosomes 1,2,3,4,5,6 and mitochondrial DNA are the same as on NCBI assembly "dicty_2.7"). Regions [3016085, 3768655] from chr2, [64985, 72996] from chrBF and [42801, 78150] from chrR were masked.
Supplementary_files_format_and_content: Processed data files are tab-separated files with two columns: the first containing gene names and the second its expression. Files ending with "_rc.tab" contain raw expression values while files ending with "_nor.tab" contain normalized expression values.
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| Submission date |
Mar 09, 2021 |
| Last update date |
Jun 15, 2021 |
| Contact name |
Gad Shaulsky |
| E-mail(s) |
gadi@bcm.edu
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| Organization name |
Baylor College of Medicine
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| Department |
Molecular and Human Genetics
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| Street address |
One Baylor Plaza
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| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
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| Platform ID |
GPL9379 |
| Series (1) |
| GSE152851 |
Transcriptional milestones in Dictyostelium development |
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| Relations |
| Reanalysis of |
GSM1517239 |
| BioSample |
SAMN18227871 |
| SRA |
SRX10292928 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSM5145730_AX4_FD_r1_hr24_nor.tab.gz |
113.2 Kb |
(ftp)(http) |
TAB |
| GSM5145730_AX4_FD_r1_hr24_rc.tab.gz |
49.1 Kb |
(ftp)(http) |
TAB |
SRA Run Selector |
| Processed data provided as supplementary file |
| Raw data are available in SRA |
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