Journal: Gene expression patterns : GEP
Article Title: Screen for Slit/Robo signaling in trunk neural cells reveals new players
Figure Lengend Snippet: Microarray of Slit2 produced log ratios, expression and classification of genes. ( A ) Cartoon of a chicken embryo electroporation. In green we show the putative plasmid solution with fast green being injected inside the neural tube. And the ± shows the position of the electrodes when giving them the electric pulses necessary for the plasmid to enter cells in the NT. ( B ) Workflow and sample preparation and array processing. For the microarray we processed 6 Total RNA samples (1 Control, 2 experimental in duplicates) for gene expression. RNA quality and concentration was determined on the Agilent Bioanalyzer and NanoDrop spectrophotometer. One-color labeling reactions were prepared using the Agilent Two-Color Protocol (Version 6.6) with 400 ng total RNA input. ( C ) Log ratio values collected after microarray show significantly upregulated genes (red), significantly down regulated genes (green), and not differentially expressed (yellow). Control samples were labeled with Cy5 and Experimental sample labeled with Cy3. RNA Spike A mix was used for Cy3 (experimental) samples and RNA spike B mix was used for all Cy5 (Control) samples and scanned at 3 μM resolution on an Agilent G2565CA High Resolution Scanner. ( D ) Percentage of gene expression upon Slit2 gain of function in microarray. The bar graphed represents 37.3% down-regulation genes and 62.7% of incidents generated up-regulated genes of NCCs Slit2 GOF vs control.
Article Snippet: Next, the labeled cRNA is fragmented and placed on the chicken 4×44K Gene Expression microarray and hybridized at 65 °C for ~17 h. Finally, the arrays are washed and scanned at 3 μM resolution on an Agilent G2565CA High Resolution Scanner.
Techniques: Microarray, Produced, Expressing, Electroporation, Plasmid Preparation, Injection, Sample Prep, Concentration Assay, Spectrophotometry, Labeling, Generated