|
| Status |
Public on Dec 30, 2017 |
| Title |
Non pregnant Fetal cotyledon_rep4 |
| Sample type |
RNA |
| |
|
| Source name |
Fetal cotyledon tissue non pregnant rep4
|
| Organism |
Bos taurus |
| Characteristics |
tissue: Fetal cotyledon
Sex: female
pregnant: no
|
| Treatment protocol |
The Gravid uteri tissue samples from pregnant buffalo were collected from local slaughter house. The samples were collected from the slaughtered animals and no animal was slaughtered purposely for tissue collection.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was perforemd by using Qiagen Rneasy mini kit with Dnase Treatment.
|
| Label |
Cy3
|
| Label protocol |
The samples for Gene expression were labeled using Agilent Quick-Amp labeling Kit (p/n5190-0442). 500ng each of total RNA were reverse transcribed at 40°C using oligo dT primer tagged to a T7 polymerase promoter and converted to double stranded cDNA. Synthesized double stranded cDNA were used as template for cRNA generation. cRNA was generated by in vitro transcription and the dye Cy3 CTP(Agilent) was incorporated during this step. The cDNA synthesis and in vitro transcription steps were carried out at 40°C. Labeled cRNA was cleaned up using Qiagen RNeasy columns (Qiagen, Cat No: 74106) and quality assessed for yields and specific activity using the Nanodrop ND-1000.
|
| |
|
| Hybridization protocol |
600ng of labeled cRNA sample were fragmented at 60ºC and hybridized on to Agilent's Bovine Gene Expression 8x15k Array (AMADID: 029411). Fragmentation of labeled cRNA and hybridization were done using the Gene Expression Hybridization kit (Agilent Technologies, In situ Hybridization kit, Part Number 5190-0404). Hybridization was carried out in Agilent’s Surehyb Chambers at 65º C for 16 hours.
|
| Scan protocol |
Agilent Microarray Scanner (Agilent Technologies, Part Number G2600D).
|
| Description |
Gene expression before pregnancy rep4
|
| Data processing |
Images were quantified using Feature Extraction Software ( Agilent). Feature extracted raw data was analyzed using GeneSpring GX software from Agilent. Normalization of the data was done in GeneSpring GX using the percentile shift normalization method .
|
| |
|
| Submission date |
Jan 19, 2017 |
| Last update date |
Jan 04, 2019 |
| Contact name |
Syed Azmal Ali |
| E-mail(s) |
azmal786@gmail.com
|
| Phone |
9728957861
|
| Organization name |
National Dairy Research Institute
|
| Department |
Animal Biotechnology
|
| Lab |
Cell biology and proteomics
|
| Street address |
G.T. Road, ABTC, Proteomics and Cell Biology Lab
|
| City |
Karnal |
| State/province |
Haryana |
| ZIP/Postal code |
132001 |
| Country |
India |
| |
|
| Platform ID |
GPL22954 |
| Series (1) |
| GSE93874 |
Genome wide gene expression analysis of differential fetal cotyledons |
|
