|
| Status |
Public on May 31, 2012 |
| Title |
fetalcellsinlung_4 |
| Sample type |
RNA |
| |
|
| Source name |
Fetal cells flow sorted from the maternal lung
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
gestational age of pups: e14
|
| Growth protocol |
Male mice homozygous for the enhanced green fluorescent protein (Egfp) transgene (C57BL/6-Tg(CAG-EGFP)C14-Y01-FM131Osb [stock number 267, Riken BioResource Center, Japan, originally provided by Dr. Masaru Okabe, bred in-house]) were mated to 10-12 week-old wild type C57BL/6J females (stock no. 664, Jackson Labs, Bar Harbor, Maine). With the homozygous male, all pups inherit one copy of the Egfp transgene. Egfp expression was used as a marker for all fetal cells independent of gender.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
GFP+, PI- fetal cells from each set of maternal lungs were sorted directly into the lysis buffer provided with the WT-Ovation One-Direct Amplification system (NuGEN Technologies, Inc., San Carlos, California) at a dilution of 10-40 cells/µl. Cell lysates from each female were pipetted repeatedly and stored at -80°C for 1-8 days following sorting. cDNA was converted and amplified from the RNA in the cell lysates using the WT-Ovation One Direct Amplification system following the manufacturer’s protocol with the exception of using up to 80 cells as input material.
|
| Label |
biotin
|
| Label protocol |
Twenty-five μg cDNA were fragmented and biotinylated using the Encore Biotin Module (NuGEN Techonologies, Inc.)
|
| |
|
| Hybridization protocol |
The arrays were hybridized in an Affymetrix oven at 45°C and 60rpm for 18-40 hours. Arrays were washed using a GeneChip Fluidics Station 450 and stained with streptavidin–phycoerythrin according to the protocol supplied by NuGEN Technologies, Inc. with their Encore Biotin Module.
|
| Scan protocol |
Arrays were scanned with the GeneArray Scanner (Affymetrix).
|
| Description |
Gene expression data from fetal cells collected from the maternal lung in the late stages of pregnancy
|
| Data processing |
The arrays were analyzed using the GeneChip Microarray Suite 5.0 (Affymetrix) and normalized in R using quantile-normalization, with ideal mismatch background correction and Tukey biweight summarization.
|
| |
|
| Submission date |
May 23, 2012 |
| Last update date |
Jun 01, 2012 |
| Contact name |
Stephanie Pritchard |
| E-mail(s) |
[email protected]
|
| Phone |
617-636-9120
|
| Organization name |
Tufts University
|
| Department |
Genetics
|
| Lab |
Diana W. Bianchi Lab
|
| Street address |
800 Washington St, Box 396
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02111 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE38188 |
Comprehensive Analysis of Genes Expressed by Rare Microchimeric Fetal Cells in Maternal Mouse Lung |
|
