ISOLATE II FFPE RNA/DNA Kit is specially designed for the rapid, phenol-free, sequential isolation of pure total RNA (including small RNA) and genomic DNA from challenging FFPE tissue samples. Formalin fixation of tissues results in crosslinking of the nucleic acids and proteins. Furthermore, the embedding process can also lead to fragmented nucleic acids with time, often resulting in lower molecular weight nucleic acids than those obtained from fresh or frozen samples. ISOLATE II FFPE RNA/DNA Kit has been developed to allow for partial reversing of formalin modifications resulting in high quality and yield of nucleic acids.
The recovery of total RNA and genomic DNA from the same sample provides the distinct advantage of providing matched nucleic acid fractions, from the same population of cells. Matched RNA/DNA is invaluable for integrative studies of the genome, epigenome and transcriptome. Maximizing RNA and DNA retrieval from a single specimen is also crucial when using tissues that are of limited availability.
Isolation is based on a fast spin column format using a novel affinity resin as the separation matrix. The RNA and DNA are preferentially purified from other cellular components without the use of phenol or chloroform, resulting in a high quality and yield of nucleic acids. The ISOLATE II FFPE RNA/DNA Kit also overcomes the bias seen in recovery of the small RNA fraction observed with phenol-based RNA recovery.
The purified RNA can be used in a number of downstream applications including real-time RT-PCR, microarrays and RNA-seq. The purified genomic DNA can be used in real-time PCR, next-generation sequencing, multiplex PCR (fig. 1) and bisulfite conversion for epigenetic analysis.
|ISOLATE II FFPE RNA Micro Columns||50|
|ISOLATE II FFPE DNA Micro Columns||50|
|Collection Tubes (2ml)||100|
|Elution Tubes (1.7ml)||100|
|Proteinase K (lyophilized)||2 x 12mg|
|Digestion Buffer DX||2 x 25ml|
|Lysis Buffer RX||40ml|
|Wash Buffer W1||2 x 38ml|
|RNA Elution Buffer||6ml|
|DNA Elution Buffer||15ml|
|DNase I Solution (RNase-free)||210µl|
|DNase I Reaction Buffer DRB||6ml|
|Bench Protocol Sheet||1|
The DNase I Solution and Proteinase K should be stored at -20°C upon arrival. All other components should be stored dry and at room temperature.
When stored under the recommended conditions and handled correctly, full activity of reagents is retained until the expiry date indicated on the outer box label.