|
| Status |
Public on Mar 18, 2016 |
| Title |
Wild-type Replicate 2 vs Brdt KO Replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
pooled Wild-type pituitary tissue
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57BL/6
genotype/variation: wild type
gender: male
age: 8 weeks
tissue: pituitary
|
| Growth protocol |
Heterozygous Brdt tm1a (EUCOMM) Wtsi mice (C57BL/6) were purchased from the International Knockout Mouse Consortium, Welcome Trust Sanger Institute, UK. The ‘knockout-first’ allele (tm1a) contains an IRES: lacZ trapping cassette and a floxed promoter-driven neo cassette inserted into the fourth intron of the Brdt gene and disrupting its function. Mice were housed and maintained in plastic cages in the animal facility at Tel Aviv Sourasky Medical Center, in a temperature-controlled room (21-25oC) with a 12-h light/12-h dark cycle.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was performed using the Trizole reagent according to the manufacturer’s instructions (Sigma, St. Louis, MO). RNA purity and integrity were analyzed by Agilent Bioanalyzer 2100 (Agilent, Santa Clara, CA).
|
| Label |
Cy5
|
| Label protocol |
Total RNA was amplified and labeled with Cy3 or Cy5 using the Low Input Quick Amp Labeling Kit, Two-Color (Agilent), following the manufacturer's instructions.
|
| |
|
| Channel 2 |
| Source name |
pooled Brdt KO pituitary tissue
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57BL/6
genotype/variation: Brdt-/-
age: 8 weeks
tissue: pituitary
|
| Growth protocol |
Heterozygous Brdt tm1a (EUCOMM) Wtsi mice (C57BL/6) were purchased from the International Knockout Mouse Consortium, Welcome Trust Sanger Institute, UK. The ‘knockout-first’ allele (tm1a) contains an IRES: lacZ trapping cassette and a floxed promoter-driven neo cassette inserted into the fourth intron of the Brdt gene and disrupting its function. Mice were housed and maintained in plastic cages in the animal facility at Tel Aviv Sourasky Medical Center, in a temperature-controlled room (21-25oC) with a 12-h light/12-h dark cycle.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was performed using the Trizole reagent according to the manufacturer’s instructions (Sigma, St. Louis, MO). RNA purity and integrity were analyzed by Agilent Bioanalyzer 2100 (Agilent, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
Total RNA was amplified and labeled with Cy3 or Cy5 using the Low Input Quick Amp Labeling Kit, Two-Color (Agilent), following the manufacturer's instructions.
|
| |
|
| |
| Hybridization protocol |
The hybridization was performed with canine-specific AMADID Release GE 4x44K microarrays (Agilent), using a Gene Expression Hybridization Kit in a hybridization oven, according to the manufacturer's instructions.
|
| Scan protocol |
Scanned on an Agilent G2565BA scanner.
|
| Description |
Wild-type Replicate 2 vs Brdt KO Replicate 2
|
| Data processing |
Raw data were normalized by Quantile algorithm, Gene Spring Software 11.0 (Agilent)
|
| |
|
| Submission date |
Sep 08, 2015 |
| Last update date |
Mar 19, 2016 |
| Contact name |
Shimi Barda |
| E-mail(s) |
[email protected]
|
| Phone |
+972524418686
|
| Organization name |
Tel Aviv Sourasky Medical Center
|
| Department |
Lis Maternity Hospital
|
| Lab |
Institute for the Study of Fertility
|
| Street address |
6 Weizman Street
|
| City |
Tel Aviv |
| ZIP/Postal code |
6423906 |
| Country |
Israel |
| |
|
| Platform ID |
GPL20889 |
| Series (1) |
| GSE72786 |
Brdt KO mice: Wild-type vs. Mutant |
|
