|
| Status |
Public on Apr 21, 2023 |
| Title |
genomic DNA from Blood from mouse 280625N at time point 2 |
| Sample type |
genomic |
| |
|
| Source name |
Blood from mouse 280625N at time point 2
|
| Organism |
Mus musculus |
| Characteristics |
age: 0.257548359
Sex: female
time point: 2
pregnancy_status: Not pregnant
mouse_id: 280625N
mousestrain: C57Bl/6J
tissue: Whole blood
|
| Treatment protocol |
C57Bl/6 mice (11 weeks old) were obtained from Jackson Laboratories. Three days before mating, male mice were separated into individual cages and soiled bedding from male cages was added to female cages to induce estrus (Whitten, 1956). 1:1 mating pairs were set up in the evening and left overnight. Females were removed from male cages the following morning and inspected for copulatory plugs. Pregnant females were identified by daily tracking of body weight. Blood was collected in EDTA tubes by submandibular vein puncture every two weeks to create a series of four samples per mouse: (1) 10 days before mating; (2) 4 days after mating; (3) 18 days after mating; and (4) 32 days after mating, generally corresponding to ~2 weeks postpartum. Pups were humanely euthanized shortly after birth allowing mothers to recover from pregnancy without needing to nurse. Blood was snap-frozen in liquid nitrogen immediately after collection and stored at –80°C until needed.
|
| Growth protocol |
This experiment was approved by the Mass General Brigham IACUC. C57Bl/6 mice were obtained from Jackson Laboratories and acclimated to our animal facility for at least 48 h before being subjected to any experimental manipulation. Mice were maintained in a barrier facility in sterilized, ventilated cages and fed standard laboratory chow (LabDiet 5053) and reverse osmosis drinking water ad libitum and maintained on a 12h:12h light:dark cycle. Mice were generally housed socially (5 mice/cage) except during the pregnancy studies wherein male mice were housed individually after mating. Mice were humanely euthanized at the conclusion of each experiment by CO2 exposure followed by cervical dislocation.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
DNA was isolated from whole blood using the DNeasy Blood and Tissue Kit (Qiagen) following the manufacturer’s protocol.
|
| Label |
Cy3,Cy5
|
| Label protocol |
Standard Illumina protocol
|
| |
|
| Hybridization protocol |
Standard Illumina protocol
|
| Scan protocol |
Standard Illumina protocol
|
| Description |
Not pregnant
|
| Data processing |
For mammalian microarray analysis, raw methylation data were first normalized and beta values were calculated using the SeSAMe R package .
|
| |
|
| Submission date |
Feb 02, 2023 |
| Last update date |
Apr 21, 2023 |
| Contact name |
Jesse Poganik |
| Organization name |
Brigham and Women's Hospital, Harvard Medical School
|
| Department |
Medicine/Genetics
|
| Street address |
77 Avenue Louis Pasteur
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
|
| Platform ID |
GPL28271 |
| Series (1) |
| GSE224352 |
Longitudinal study of DNA methylation changes during mouse pregnancy |
|
