|
| Status |
Public on Dec 15, 2009 |
| Title |
mRNA_delayed_digital_gene_expression |
| Sample type |
SRA |
| |
|
| Source name |
delayed_implantation
|
| Organism |
Mus musculus |
| Characteristics |
tissue: uterus
|
| Treatment protocol |
Female mice were mated with fertile males of the same strain to induce pregnancy (day 1 is the day of vaginal plug). To induce delayed implantation, pregnant mice were ovariectomized under ether anesthesia at 08:30-09:00 h on day 4 of pregnancy. Delayed implantation was maintained from days 5-7 by injecting progesterone (1 mg/mouse, Sigma). Estradiol-17 (25 ng/mouse, Sigma) was given to progesterone-primed delayed implantation mice to initiate implantation on day 7 of pregnancy. The mice were sacrificed to collect uteri 24 h after estrogen treatment for activation group. Delayed implantation was confirmed by flushing the blastocysts from one horn of the uterus. The implantation sites of activated uterus were identified through intravenous injection of 0.1 ml of 1% Chicago blue.
|
| Growth protocol |
Mature mice (Kunming White outbred strain) were maintained in a controlled environment with a 14-h light/10-h dark cycle. All animal procedures were approved by the Institutional Animal Care and Use Committee of Xiamen University.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
mRNAs were isolated through binding to a magnetic oligo(dT) beads. First strand cDNA was synthesized using the binding mRNA as a template, and followed by the synthesis of the second strand of cDNA. After cDNAs were digested with DpnII, the double stranded cDNA fragment attached to the oligo(dT) bead was collected and ligated to a GEX DpnII adapter 1 at the site of DpnII cleavage. The sequence for MmeI was included in GEX DpnII adapter 1. After purification, products coupled with oligo(dT) bead were digested with MmeI to create a 16 bp tag, followed by ligating to a GEX adapter 2 at the site of MmeI cleavage. After PCR amplification, the purified DNA fragments were used directly for sequencing using the Illumina Cluster Station in Shenzhen Huada Gene Sci-Tech Company.
polyA+ mRNA by oligo dT beads
|
| |
|
| Library strategy |
RNA-Seq |
| Library source |
transcriptomic |
| Library selection |
cDNA |
| Instrument model |
Illumina Genome Analyzer II |
| |
|
| Description |
mRNA tag profiling (digital gene expression) analysis of delayed implantation
|
| Data processing |
processed data build: mm9, the 3’ end of the read was determined by the 3’ most perfect match to the first 8 nt of the 3’ adaptor.
|
| |
|
| Submission date |
Dec 15, 2009 |
| Last update date |
May 15, 2019 |
| Contact name |
Zeng-Ming Yang |
| E-mail(s) |
zmyang@stu.edu.cn
|
| Phone |
0754-82902011
|
| Organization name |
Shantou University
|
| Department |
Biology
|
| Lab |
Reproductive Biology
|
| Street address |
263 Daxue Road
|
| City |
Shantou |
| ZIP/Postal code |
515063 |
| Country |
China |
| |
|
| Platform ID |
GPL9250 |
| Series (1) |
| GSE19473 |
The integrative analysis of microRNA and mRNA expression in mouse uterus under delayed implantation and activation |
|
| Relations |
| SRA |
SRX015123 |
| BioSample |
SAMN00007207 |
