GDP439-RNAprep Pure FFPE Kit

RNAprep Pure FFPE Kit
For purification of total RNA from formalin-fixed, paraffin-embedded tissue sections

RNAprep Pure FFPE Kit
(Spin Column)
Cat. no. GDP439

Storage
DNase I, Buffer RDD & RNase-Free ddH2O (1 ml) should be stored at
2-8°C for 15 months; others stored at room temperature (15-30°C) for 15months.

Introduction
RNAprep Pure FFPE Kit is specially designed for purification of total RNA
from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Due to
fixation and embedding conditions, nucleic acids in FFPE samples are
usually heavily fragmented and chemically modified by formaldehyde.
Therefore, nucleic acids isolated from FFPE samples is often of a lower
molecular weight than those obtained from fresh or frozen samples. RNA
obtained by this kit could be used for downstream experiment as RT-PCR.

Important Notes

1. Add ethanol (96-100%) to Buffer RW before start as indicated on the
   bottle.
2. Preparation of DNase I stock solution
   Dissolve the lyophilized DNase I (1500 U) in 550 μl of the RNase-Free
   ddH2
   O. Mix gently by inverting. Do not vortex. Divide it into single-use
   aliquots, and store at -30~-15°C for up to 9 months.
   Note: Thawed aliquots from -30~-15°C could be stored at 2-8°C for up
   to 6 weeks. Do not refreeze the aliquots after thawing.

Starting Material

1. Standard formalin-fixation and paraffin-embedding procedures always
   result in significant fragmentation and crosslinking of nucleic acids. To
   limit the extent of nucleic acid fragmentation and crosslinking, be sure
   to:

• Fixate tissue samples in 4-10% formalin buffer as quickly as possible
  after surgical removal;
• Use a fixation time of 14-24 hours (longer fixation time leads to overfixation and more severe nucleic acid fragmentation, resulting in poor
  performance in downstream assays);
• Thoroughly dehydrate samples prior to embedding.

2. The starting material for RNA purification should be freshly cut sections
   of FFPE tissue, each with a thickness of up to 10 μm. Thicker sections
   may result in lower nucleic acid yields. Up to 8 sections, each with a
   thickness of up to 10 μm and a surface area of up to 250 mm
   2, can be combined in one preparation.
3. If there is no information about the nature of your starting material,
   we recommend starting with no more than 2 sections per preparation.Depending on RNA yield and purity, it may be possible to use up to 8 sections in subsequent preparations.

GDP439-RNAprep Pure FFPE Kit

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