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The Mag-Bind® FFPE RNA 96 kit provide a rapid and easy method for the isolation of
total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Due to fixation
and embedding procedures, nucleic acids in FFPE samples are heavily fragmented and
modified by formaldehyde. While the Mag-Bind® FFPE RNA 96 kit are optimized to
minimize the effect of the formaldehyde modification, it is not recommended to use the
RNA purified with this kit for downstream applications that requires full length RNA.
The Mag-Bind® FFPE RNA 96 kit combine high efficient binding properties of Mag-Bind®
technology with a specially designed buffer system to isolate total RNA sample from
FFPE sample. There are two protocols included in this manual. The standard protocol
uses a heating step instead of xylene to remove paraffin from the sample. The alternative
protocol uses traditional xylene extraction to remove paraffin.
Samples are first lysed in RML Buffer with digestion of Proteinase K. The lysate is then
mixed with MFB Buffer and magnetic particles to bind the nucleic acid on the surface of
the Mag-Bind® Particles SC. Genomic DNA is removed by DNase I digestion. After two
wash steps, purified RNA is eluted with RNase-free water.
Since standard formalin fixation and paraffin embedding procedures cause significant
fragmentation of nucleic acids. We recommend following guidelines to limit the extent
of DNA/RNA fragmentation: 1) Use 4-10% formalin to fixate tissue samples; 2) Limit
the fixation time to 14-24 hours; 3) Completely dehydrate samples before embedding.
Always use freshly cut sections of FFPE tissue for RNA isolation. For the first time user, we
recommend to use less than 3-5 sections with thickness of 10 μm. Depending on the yield
and purity obtained, it may be possible to increase the starting material.
New in this Edition:
March 2018
•
The storage temperature for DNase I Digestion Buffer has changed. DNase I Digestion
Buffer should now be stored at -20°C along with the Mag-Bind® DNase I.
•
The 200 prep size of M2551, Mag-Bind® FFPE RNA 96 Kit, has been discontinued and
is no longer available to purchase.
•
M2555, Mag-Bind® FFPE RNA Kit, has been discontinued and is no longer available to
purchase.
January 2014
•
DNase I has been renamed Mag-Bind® DNase I. This is a name change only.
•
Proteinase K is now supplied in a liquid form eliminating the step to resuspend prior
to use. Proteinase K Solution can also be stored at room temperature for 12 months.
•
Proteinase Storage Buffer is no longer included in the kit.
2
Introduction
Principle
Starting Materials
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