|
| Status |
Public on Feb 26, 2008 |
| Title |
FoxH1 morpholino-injected 40% epiboly-stage embryos vs. control morpholino-injected 40% epiboly-stage embryos_ZF1221 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source Name |
control morpholino-injected 40% epiboly-stage zebrafish embryos
|
| Organism(s) |
Danio rerio |
| Characteristics |
Zebrafish embryos
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with Trizol/Chloroform method according to manifecturer protocol and the RNA is further purified by the Qiagen RNAeazy columns
|
| Label |
Cy5
|
| Label protocol |
10-15 microgram of total RNA was reverse transcribed with Anchor Oligo-dT and Aminoallyl dUTP, after NaOH RNA hydrolysis, the labeled Aminoallyl dUTP cDNA is purified with Qiagen, QIAQuick PCR Purification Kit (Cat. # 28104). The Aminoallyl dUTP cDNAs are then coupled with either Cy3 or Cy5 mono NHS Ester Dyes. The CyDyes labeled cDNAs are then purified with Qiagen, QIAQuick PCR Purification Kit (Cat. # 28104).
|
| |
|
| Channel 2 |
| Source Name |
FoxH1 morpholino-injected 40% epiboly-stage zebrafish embryos
|
| Organism(s) |
Danio rerio |
| Characteristics |
Zebrafish embryos
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted with Trizol/Chloroform method according to manifecturer protocol and the RNA is further purified by the Qiagen RNAeazy columns
|
| Label |
Cy3
|
| Label protocol |
10-15 microgram of total RNA was reverse transcribed with Anchor Oligo-dT and Aminoallyl dUTP, after NaOH RNA hydrolysis, the labeled Aminoallyl dUTP cDNA is purified with Qiagen, QIAQuick PCR Purification Kit (Cat. # 28104). The Aminoallyl dUTP cDNAs are then coupled with either Cy3 or Cy5 mono NHS Ester Dyes. The CyDyes labeled cDNAs are then purified with Qiagen, QIAQuick PCR Purification Kit (Cat. # 28104).
|
| |
|
| |
| Hybridization protocol |
The combined Cy3 and Cy5 labeled cDNAs are put in a total volume of 250 ul DEPC H20 and 250 ul of 2X Agilent hybridization buffer. The mixture is then denatured for 2 minutes and applied on the Agillent hybridization chamber/gasket slide in sandwich with the array slide. The hybridization is performed at 65 degre for 16 hours.
|
| Scan protocol |
Agilent Scanner
|
| Description |
Zebrafish embryos
|
| Data processing |
IPLab/DeArray
|
| |
|
| Submission date |
Feb 26, 2007 |
| Contact name |
Christopher Pan |
| E-mail(s) |
cpan@mail.nih.gov
|
| Organization name |
NIH
|
| Department |
NHGRI
|
| Street address |
Rm 5228, Bldg 50, South Dr.
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
| |
|
| Platform ID |
GPL4609 |
| Series (1) |
| GSE8654 |
Identification of mesendoderm precursor- and neurectoderm precursor-enriched genes in Zebrafish embryos |
|
