Takara NucleoSpin totalRNA FFPE XS, 50 Preps

The NucleoSpin totalRNA FFPE kit (50 preps) allows for the purification of small and large RNA species from very small FFPE samples (<5 mg tissue) into a 5–30 µl elution volume. The kit includes NucleoSpin RNA FFPE XS Columns, Collection Tubes, Paraffin Dissolver, buffers, Proteinase K, and RNase-free rDNase.

NucleoSpin XS columns are designed for efficient elution of DNA/RNA in small volumes.Panel A. The columns are optimized to hold a very small silica membrane, yet fit inside a standard 1.5 mL microfuge tube.Panel B. The exceptionally small (2.0 mm) diameter of the NucleoSpinXS membrane (Column MN) allows efficient elution in only 5–30 μl. It provides a significant advantage over the small-sample column offered by Competitor Q (Column Q; 4.8 mm diameter), which requires an elution volume of at least 10–30 μl.

NucleoSpin totalRNA FFPE provides better RT-PCR performance (lower CTvalues) than competitor kits, indicating higher RNA yields.Large (e.g., mRNA) and small (e.g., miRNA) RNA were isolated from 4 x 10 µm FFPE sections of mouse brain tissue with NucleoSpin totalRNA FFPE and three competitor kits (Q, A, P). Quantification of mRNA* and miRNA** was performed using qRT-PCR. Low CTvalues indicate high RNA yields.*The amplification target consists of a 230-base fragment of the β2-microglobulin gene.**miRNA was quantified using the miR-16 MicroRNA Assay in the TaqMan MicroRNA Reverse Transcription Kit from Applied Biosystems.

NucleoSpin totalRNA FFPE provides more efficient gDNA removal than competitor kits.Large (e.g., mRNA) and small (e.g., miRNA) RNA were isolated from 4 x 10 µm FFPE sections of mouse brain tissue with NucleoSpin totalRNA FFPE and three competitor kits (Q, A, P). Residual DNA was assayed by amplifying a 191-bp fragment of the mGAPDH gene. Higher CTvalues correspond to lower amounts of residual DNA.

The higher concentration of total RNA obtained from XS columns allows efficient, sensitive qRT-PCR from very small FFPE samples.Trace amounts of FFPE samples (1 x 15 µm mouse spleen and 1 x 10 µm mouse heart) were used to isolate mRNA and miRNA with NucleoSpin totalRNA FFPE and NucleoSpin totalRNA FFPE XS. The RNA was eluted in 50 μl (using a NucleoSpin totalRNA FFPE mini spin column) and 20 μl (using a NucleoSpin totalRNA FFPE XS column). The mRNA* and miRNA** eluted from the XS column at higher concentrations, reducing the CTvalue by at least one cycle. When using standard FFPE sample amounts, NucleoSpin totalRNA FFPE (which uses mini spin columns) is recommended in order to avoid possible column overloading.*The amplification target consists of a 230-base fragment of the β2-microglobulin gene.**miRNA was quantified using the miR-16 MicroRNA Assay in the TaqMan MicroRNA Reverse Transcription Kit from Applied Biosystems