Suppression subtractive hybridization profiles of radial growth phase and metastatic melanoma cell lines reveal novel potential targets-2

Decorin) (), () and () of the RGP library; (14-3-3 ξ) () identified in both libraries; and () and () from the Met library were isolated and used as probes for hybridization in Northern blots containing total RNA from the melanoma cell lines indicated above the panels – mean that the corresponding cell line was not included in the Northern blot, and were introduced to allow alignment among panels. Northern blots were prepared as described in Figure 1. () identified in the Met library was validated by RT-PCR. For RT-PCR, total RNA samples (2 μg) from the indicated cell lines were, after DNase treatment, submitted to reverse transcription with Superscript II (Invitrogen) using oligo dT as primer and the cDNA was used as template for PCR amplification with primers. After 25, 28, 30 and 32 amplification cycles, 5 μl aliquots were collected for agarose gel electrophoresis. As endogenous control, a pair of primers for the (β-actin) mRNA was used. : Control RT-PCR amplification using as template RNA (DNase treated) without prior reverse transcription.<p><b>Copyright information:</b></p><p>Taken from "Suppression subtractive hybridization profiles of radial growth phase and metastatic melanoma cell lines reveal novel potential targets"</p><p>http://www.biomedcentral.com/1471-2407/8/19</p><p>BMC Cancer 2008;8():19-19.</p><p>Published online 22 Jan 2008</p><p>PMCID:PMC2267200.</p><p>