Total Nucleic Acid Isolation for FFPE Samples

FFPE (formalin-fixed paraffin-embedded) samples are an important resource for the discovery and validation of cancer biomarkers. Millions of archived samples are available worldwide for retrospective studies; however, there are significant challenges associated with these types of samples—especially for gene expression analysis— due to RNA degradation, chemical modification, and limited sample quantity. In addition, differences in fixation and RNA isolation protocols as well as sample age further affect data quality.
UTSW Microarray Core Facility is providing service for the extraction of total nucleic acid from formalin- or paraformalin-fixed, paraffin-embedded (FFPE) tissues. Recovered nucleic acids are suitable for next generation sequencing, real-time RT-PCR, PCR, mutation screening, and microarray analyses
Sample preparation :
If you are preparing tissue sections, cut 5–20 μm sections from FFPE tissue blocks using a microtome. Place the equivalent of ≤ 80 μm of tissue slices (i.e., a maximum of four 20 μm, eight 10 μm, or sixteen 5 μm slices) in a 1.5 mL microcentrifuge tube. (Note: For recovery of miRNA, we recommend using ≥10 μm slices).
If you are preparing unsectioned core samples, cut an unsectioned core sample, up to 35 mg, from a paraffin block. Crush or grind the sample in liquid nitrogen or on dry ice, and place in a 1.5 mL microcentrifuge tube.