|
| Status |
Public on Jan 31, 2008 |
| Title |
Embryonic liver age 28 IOF replica 2 |
| Sample type |
RNA |
| |
|
| Source name |
Total RNA from liver enbryos day 28 labeled with cyanine-3 (green)
|
| Organism |
Meleagris gallopavo |
| Characteristics |
Tissue: liver
28E IOF
|
| Treatment protocol |
16 eggs were sampled at each age (22, 24, 25, 26 and 28 days of incubation) to harvest the liver; tissue preserved in RNA Later solution (Ambion, Inc). In ovo injection was performed on day 24 of incuibation. Samples from days 25, 26 and 28 of incubation were collected for both: control and in ovo fed embryos.
|
| Growth protocol |
turkey eggs incubated under standard conditions
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions (Molecular Research Center, Inc.)
|
| Label |
Cy3
|
| Label protocol |
5µg of total RNA were primed with 3µg of random primers and 2µg of oligo(dt) primer, heated at 70C for 10 min, then reverse transcribed at 42C for 2h with 3.2 µl ChipShot Reverse Transcriptase and 4 µl aminoallyl-sNTPs; cDNA was indirect labeled with CyDye NHS Esters (Cy3 or Cy5) by adding 60 µl of cDNA eluted to dried 40,000 pmol CyDye aliquot and hybrydized at room temperature for 1h, followed by purification steps as indicated by manufacturer's instructions (ChipShot Indirect Labeling System and Membrane Clean-Up System, Promega Corp)
|
| |
|
| Hybridization protocol |
mix of 33 pmol of both samples was resuspended with 33 µl Pronto! Long Oligo Hybridization solution and applied to pre-hybrydized microarray slides with spotted area coved with a 25 X 40 mm glass coverslip; slides were hybridized in Corning Hybridization Chambers immersed in 42C water bath, protected from light, for 16h. After hybridization, slides were sequencenly washed according to manufacturer's instructions (Pronto! Universal Microarray Hybridization Kit, Corning Inc.)
|
| Scan protocol |
scanned on ScanArray Gx Plus (PerkinElmer) set to 65% lazer power
Images were quantified using ScanAlyze Software (version 2.50).
|
| Description |
Slide 11-Embryonic liver: age 28 IOF replica 2 vs age 26 IOF replica 3
|
| Data processing |
Log2 transformed data was submitted to ratio analysis and LOWESS normalization using JMP Genomics Software (SAS Institute)
|
| |
|
| Submission date |
Oct 23, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Jean Eduardo de Oliveira |
| Phone |
919-515-5537
|
| Fax |
919-515-7070
|
| URL |
http://www.ncsu.edu/ashwell-lab/
|
| Organization name |
North Carolina State University
|
| Department |
Poultry Science
|
| Lab |
Nutrition/Domestic Animal Genomics
|
| Street address |
2711 Founders Dr
|
| City |
Raleigh |
| State/province |
NC |
| ZIP/Postal code |
27695-7608 |
| Country |
USA |
| |
|
| Platform ID |
GPL6043 |
| Series (2) |
| GSE9410 |
Effect of in ovo feeding on late-term turkey embryo liver gene expression profiling |
| GSE9532 |
Effect of in ovo feeding on late-term turkey embryo in 5 tissues |
|
