|
| Status |
Public on Dec 31, 2007 |
| Title |
stage13_mef2(424)_Rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Whole embryo Stage 13 mef2(424) Allele
|
| Organism |
Drosophila melanogaster |
| Characteristics |
Age: Stage 13, Tissue: Whole embryo
|
| Treatment protocol |
The staged, dechorionated stage 13 embryos were immediately homogenised in TriZol prior to storage at –80ºC.
|
| Growth protocol |
Flies were raised in standard conditions. Embryo collections were as follows. Following 2 x 1 hour prelays, 30 minute embryo collections were aged at 25ºC for 6.5 hours prior to sorting. Mutant embryos were individually selected by absence of the GFP fluorescent balancer chromosome (CyO P(Gal4-twi)P(UAS-2eGFP)). Both mutant and control embryos were accurately staged at mid 12 using autofluorescence to visualise the germ band to ensure that all were within a 30-minute window. Pools of 150 embryos were allowed to develop until mid stage 13, dechorionated, inspected to ensure normal development had continued.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TriZol (Invitrogen) and cleaned up by RNeasy mini-kit (Qiagen) according to the manufacturer's instructions
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the standard Affymetrix protocol from 5 ug total RNA (Expression Analysis Technical Manual, 2004, Affymetrix)
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA was hybridized for 16 hr at 45ºC on Affymetrix GeneChip Drosophila Genome 1 Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip scanner 3000 7G.
|
| Description |
Drosophila_embryo_stage13_mef2(424)_Rep2
|
| Data processing |
Data were processed and normalised using MAS5.0 algorithm
|
| |
|
| Submission date |
Dec 14, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Michael Victor Taylor |
| E-mail(s) |
TaylorMV@cardiff.ac.uk
|
| Phone |
+44 2920875881
|
| URL |
http://www.cf.ac.uk/biosi/research/genetics/staff/taylor.html
|
| Organization name |
Cardiff University
|
| Department |
School of Biosciences
|
| Lab |
Taylor
|
| Street address |
Main Building, Park Place
|
| City |
Cardiff |
| State/province |
S. Glam |
| ZIP/Postal code |
CF103TL |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL72 |
| Series (1) |
| GSE9889 |
mef2 activity levels differentially affect gene expression during Drosophila muscle development |
|
