|
| Status |
Public on Aug 30, 2024 |
| Title |
TG Non-Tumor Burden Mouse 2 |
| Sample type |
RNA |
| |
|
| Source name |
Trigeminal ganglia from age-matched non-tumor burdened mouse
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6
gender: Male
tumor: None
|
| Treatment protocol |
Mice were injected with 5 x 104 in 50 uL of cell culture media. Mice were anesthetized with a ketamine (87.5 mg/kg)/xylazine (10 mg/kg) mixture for initial injection. Mice were injected orthotopically in oral cavity with a 25 gauge needle. Tumors were measured every 7 days with caliper measurements while mice were sedated with isofluorane.
|
| Growth protocol |
All animal experiments were performed in the Sanford Research Animal Resource Center which is a specific pathogen free facility. All mice were maintained in IVC Tecniplast Green line Seal Safe Plus cages. Animal rooms were maintained at 75°F, 30-70% humidity, with a minimum of 15 air changes per hour, and a 14:10 light/dark cycle. MOC2-7 cells were maintained in DMEM + 10%FBS + 1% pencillin/streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
At day 28 post injection, mice were euthanized with CO gas and desaguinated with PBS so to maintain CNS integrity. Trigeminal ganglia were dissected from the skull base and RNA was isolated via Qiazol extraction reagent and further isolated through phenol-chloroform purification.
|
| Label |
SYBR Green
|
| Label protocol |
cDNA was generated by RT reaction using High Capacity cDNA Reverse Transcription kit from Applied Biosciences. cDNA was used to run on ScienCell qPCR arrays per manufacturer and analyzed on CFX-96 BioRad.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Control
|
| Data processing |
The normalization and data analysis was conducted according to manufacturers instructions. For the normalization, five housekeeping genes are included (actinb, B2M, Gusb, Gadph, and Hrpt). For normalization, actinb was not included in normalization due to significant expression differences between test and control samples. Normalization used the averages of B2M, Gusb, Gadph, and HRPT. Target gene signals were normalized to housekeeping genes: 2^-delta(Ct), where delta(Ct) = (CT_target - CT_HKG).
|
| |
|
| Submission date |
Aug 22, 2024 |
| Last update date |
Aug 30, 2024 |
| Contact name |
Paola Drapkin Vermeer |
| E-mail(s) |
paola.vermeer@sanfordhealth.org
|
| Phone |
605 312 6110
|
| Organization name |
Sanford Research
|
| Department |
Cancer Biology and Immunotherapies Group
|
| Street address |
2301 East 60th street north
|
| City |
Sioux Falls |
| State/province |
SD |
| ZIP/Postal code |
57104 |
| Country |
USA |
| |
|
| Platform ID |
GPL34835 |
| Series (1) |
| GSE275481 |
GeneQuery RNA Profile of Trigeminal Ganglia from Tumor Burden and Tumor Naïve Mice |
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