|
| Status |
Public on Jul 06, 2014 |
| Title |
CorticalProgenitors_Fezf2_48hr_rep3 |
| Sample type |
RNA |
| |
|
| Source name |
in utero electorporated cortex, dissociated and FACS sorted
|
| Organism |
Mus musculus |
| Characteristics |
strain: CD1
expression vector: Fezf2GFP
time of purification: 48hr
|
| Treatment protocol |
800 nl of purified DNA (2 μg/μl, Fezf2GFP and CtrlGFP construct) were mixed with 0.005% Fast Green in sterile PBS and were injected in utero into the lateral ventricle of CD1 embryos at E14.5 under ultrasound guidance (Vevo 770, VisualSonics). Five 40 volt pulses of 50 ms were delivered at 1 second intervals in an appropriate orientation across the embryonic head using 1 cm-diameter platinum electrodes placed outside the uterus, using a CUY21EDIT square wave electroporator (Nepa Gene). Injected embryos were collected for FACS purification either 24 hours or 48 hours after electroporation.
|
| Growth protocol |
N/A, cells were FACS purified from in vivo sources
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Fezf2GFP and CtrlGFP electroporated cells were purified by FACS directly into RNAlater7 and RNA was extracted with the StrataPrep Total RNA Micro Kit (Stratagene), and RNA quality was assessed using a bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
RNA was amplified per Affymetrix small sample protocol, using two consecutive rounds of linear in vitro transcription to obtain 15–20 μg of amplified and labeled cRNA for each hybridization
|
| |
|
| Hybridization protocol |
Hybridization was performed according to Affymetrix protocols.
|
| Scan protocol |
Scanning was performed according to Affymetrix protocols.
|
| Data processing |
Data from individual microarrays normalized by GC-RMA. Fold changes were calculated in Rosetta Resolver
|
| |
|
| Submission date |
Apr 02, 2014 |
| Last update date |
Jul 06, 2014 |
| Contact name |
Paola Arlotta |
| Organization name |
Harvard University
|
| Department |
Stem Cell and Regenerative Biology
|
| Lab |
Paola Arlotta
|
| Street address |
7 Divinity Ave SF301-7
|
| City |
Cambridge |
| State/province |
Massachusetts |
| ZIP/Postal code |
02138 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE56451 |
Fezf2 overexpression in murine cortical progenitors in vivo |
|
