|
| Status |
Public on Mar 01, 2024 |
| Title |
Wild type Dark 2 days before fertilization replica 3 |
| Sample type |
SRA |
| |
|
| Source name |
Sexual development tissue
|
| Organism |
Neurospora crassa |
| Characteristics |
time point: 2 days before fertilization
strain: OR74A
genotype: Wild type
treatment: Dark
|
| Treatment protocol |
For this experiment, strains of complementary mating type, ∆ve-1 mat A and wild type mat a were used.. Samples were collected before the crossing (2 and 1 day prior fertilization), right before crossing (day 0) and 1, 2, 4, and 6 days after fertilization.
|
| Growth protocol |
Induction of formation of protoperithecia were carried out by spreading a total of 2x104 conidia into nitrogen-poor Synthetic Crossing Medium (SCM) agar´s plates and then incubated at 22°C with light or in darkness, depending on the experiment, for 7 days to allow the formation of protoperithecia. Conidia from a strain of opposing mating type were collected and suspended in 2.5% Tween 20. Formation of protoperithecia in each strain was examined using a stereomicroscope between days 5 to 7 prior to fertilization. Crosses were performed by applying 2 ml of a conidia suspension (5x105 conidia/ml with 2.5% Tween 20) to the surface of the protoperithecium plates. After fertilization, sexual development occurred under the same culture conditions as protoperithecium development, and all the process was monitored using stereomicroscope.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Fungal material was harvested by scrapping the surface with a razor blade. Samples were collected before the crossing (2 and 1 day prior fertilization), right before crossing (day 0) and 1, 2, 4, and 6 days after fertilization. All tissues and perithecia were immediately frozen in liquid nitrogen and stored at -80°C.
Poly-A selection and cDNA libraries preparation for RNA were prepared according to the Illumina mRNA Sample Preparation Guide
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina NovaSeq 6000 |
| |
|
| Description |
WTDtm2_R3
|
| Data processing |
libraries were run on Genome Analyzer (Illumina NovaSeq 6000).
paired-end reads of 150 nucleotide each were mapped against the N. crassa OR74A genome by using Star 2.7
quality was checked using FastQC
automatic preprocessing of the reads was carried out with AFTERQC 0.9.6
SeqMonk was used for visualization and quantification
Supplementary files format and content: raw counts, matrix text format
|
| |
|
| Submission date |
Feb 08, 2023 |
| Last update date |
Mar 01, 2024 |
| Contact name |
Gabriel Gutierrez |
| Organization name |
University of Seville
|
| Department |
Genetics
|
| Street address |
PO BOX 1095
|
| City |
Sevilla |
| ZIP/Postal code |
41080 |
| Country |
Spain |
| |
|
| Platform ID |
GPL26551 |
| Series (1) |
| GSE224796 |
Transcriptional regulation by the Velvet complex during sexual development in Neurospora crassa |
|
| Relations |
| BioSample |
SAMN33209897 |
| SRA |
SRX19312749 |
