从福尔马林固定的石蜡包埋(FFPE)模块中提取总 RNA

Zhihai Ma

doi:10.21769/BioProtoc.161

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Peer-reviewed

Total RNA Extraction from Formalin-Fixed, Paraffin-Embedded (FFPE) Blocks

从福尔马林固定的石蜡包埋(FFPE)模块中提取总 RNA

ZM Zhihai Ma email

发布: 2012年04月05日第2卷第7期 DOI: 10.21769/BioProtoc.161 浏览次数: 29682

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The authors used this protocol in:

Cover of The Journal of Molecular Diagnostics, featuring study using the protocol.

Sep 2009

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Abstract

Total RNA is extracted from fixed biological specimens by this method with higher yield than commercial kits. The product contains intact micro RNAs and small RNAs, and fragmented long RNAs.

Materials and Reagents

Paraffin embedded specimen
100% xylene (Thermo Fisher Scientific, catalog number: 6601 )
100% ethanol
Protease K (Life Technologies, Ambion^®, catalog number: AM2546 )
Trizol (Life Technologies, Invitrogen™, catalog number: 15596-018 )
Chloroform (Sigma-Aldrich, catalog number: C2432 )
Glycogen (F. Hoffmann-La Roche, catalog number: 10901393001 )
Isopropyl alcohol (Isopropanol)
RNase-free water or TE buffer (USB, catalog number: 75834 ; Promega Corporation, catalog number: P1193 )
Tris-HCl (Life Technologies, Invitrogen™, catalog number: 15568-025 )
CaCl₂ (Sigma-Aldrich, catalog number: C5670 )
Sodium dodecyl sulfate (Life Technologies, Invitrogen™, catalog number: 15525-017 )
Commercial kit (Life Technologies, Ambion^®, Austin, TX)
Sodium dodecyl sulfate
Protease digestion buffer (see Recipes)