|
| Status |
Public on Jul 01, 2011 |
| Title |
EmbryonicCleft_Age12 rep 2 |
| Sample type |
RNA |
| |
|
| Source name |
Salivary gland, replicate 2
|
| Organism |
Mus musculus |
| Characteristics |
strain: ICR
collection method: laser microdissection
age: Embryonic day E12
tissue: salivary gland
salivary gland domain: Cleft
|
| Treatment protocol |
No special treatment of pregnant mice. Embryos were isolated from timed pregnancies, the salivary glands isolated and embedded in OCT on top of nucleopore filters. The filters were removed, and the salivary glands sectioned at 10um. The sections were stained, and specific locations isolated by laser microdissection.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The RNA from the isolated cell samples were extracted using either Trizol or Rneasy Micro (Qiagen), and the RNA precipitated using glycogen as a carrier. The samples were then amplified using MessageAMP 2AA, and the spike ins incorporated at the start of the 2nd round.
|
| Label |
Cy3
|
| Label protocol |
MessageAMP 2 AA labeling protocol, however, ratio of aaUTP to UTP was reduced from 1:1 to 1:3 to reduce background signal. The RNA was then labeles following the manufacturers instructions.
|
| |
|
| Hybridization protocol |
Standard Agilent hybridization protocol: 1.5 mg of labeled RNA were fractionated and hybridized overnight (17h) at 60C, followed by washing according to Agilent's instructions.
|
| Scan protocol |
Standard Agilent scan using Agilent Microarray Scanner. Sensitivities used were 10 and 100%
|
| Description |
laser microdissected sample
E12.5Cl_002
|
| Data processing |
GeneSpring 11.0, Quantile normalization of microarray data
|
| |
|
| Submission date |
Jul 08, 2010 |
| Last update date |
Jul 01, 2011 |
| Contact name |
Kurt Musselmann |
| E-mail(s) |
[email protected]
|
| Phone |
+13017607402
|
| Organization name |
NIDCR
|
| Department |
Laboratory of Cell and Developmental Biology
|
| Lab |
Cell Biology Section
|
| Street address |
30 Convent Dr, Room 426
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL7202 |
| Series (1) |
| GSE22828 |
Atlas of Gene Expression at Microanatomical Resolution of the Submandibular Salivary Gland |
|
