|Public on Feb 22, 2011
|Xenopus laevis CNC from stage 17 embryos injected with a morpholino to ADAM13 and a GFP-fusion protein containing the ADAM13 cytoplasmic domain.
|developmental stage: Stage 15-17 embryos
tissue: cranial neural crest explant
agent: a morpholino to ADAM13 and a GFP-fusion protein containing the ADAM13 cytoplasmic domain
|Cranial neural crest cells were dissected and imediately extracted.
|Embryos were injected at the one cell stage and grown at 15C until stage 15 to 17.
|RNA extraction was performed as previously described using guanidium thioisocyanate, phenol chlorophorm and lithium chloride precipitation. P. Chomczynski, N. Sacchi, Anal. Biochem. 162, 156 (1987).
|Total RNA quality control was performed with the Bioanalyzer 2100 (Agilent). Double stranded cDNA was generated and labeled with the Ovation RNA Amplification System and Encore Biotin Module (NuGen), respectively. Purification was performed using the Qiagen PCR Purification Kit
|Labeled cDNA were incorporated into a hybridization solution from the Affymetrix GeneChip Hybridization Kit. Each sample was hybridized in separate arrays for 20 hours at 450C.
|All arrays were treated by the MIT core facility. http://openwetware.org/wiki/BioMicroCenter:Microarrays.
|10 ng of MO at the one cell stage plus 300pg of mRNA.
MO13+C13-4 (expt DA4).
|Initial data analysis was generated with the Affymetrix Expression Console and an R package was used to assess data quality. Sample normalization was done using the gcrma method.
|Apr 26, 2010
|Last update date
|Feb 22, 2011
|University of Massachusetts
|Vet & Animal Sciences
|661 North pleasant street
|ADAM13 knockdown in Xenopus laevis cranial neural crest