QIAamp DNA FFPE Tissue Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Publication protocol

Gene microarrays
For the gene microarrays, we compared three disseminated plus one appendiceal PMP samples with three samples of normal colonic mucosa. Fresh frozen samples were sectioned and stained with cresyl violet (LCM Staining Kit, Ambion, Life Technologies, Paisley, UK). Epithelial cells were captured from approximately 6 sections per sample using a Leica LMD6000 system, and to avoid any bias in cell lineage full length crypts from normal samples were captured wherever possible. RNA was isolated from each sample immediately after laser capture using a Qiagen RNAeasy plus micro kit (Qiagen, Crawley, West Sussex, UK) as per the manufacturer's instructions. The isolated RNA was quantified using an Agilent bioanalyzer and amplified using a NuGEN WT-Ovation™ RNA Amplification System (NuGEN, Leek, The Netherlands) as recommended by the manufacturer. The amplified RNA was hybridized to Affymetrix Genechip Human Exon 1.0 ST Arrays at the Cancer Research UK Affymetrix Genechip Microarray Service. Probesets were filtered to retain exonic, reliable, uniquely mapping probesets and retained probesets were also filtered according to DABG score (detection above background), to keep only those probesets reliably detected in at least two of the 7 samples.

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Papers

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Manufacturer protocol

Download the product protocol from Qiagen for QIAamp DNA FFPE Tissue Kit below.

Download manufacturer protocol

Videos

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