AllPrep DNA/RNA FFPE Kit – Nucleic Acid Extraction

For simultaneous purification of genomic DNA and total RNA (including small RNAs) from formalin-fixed, paraffin-embedded tissue sections

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AllPrep DNA/RNA FFPE Kit (50)

Cat no. / ID.   80234

50 RNeasy MinElute Spin Columns, 50 QIAamp MinElute Spin Columns, Collection Tubes, RNase-Free Reagents, and Buffers
$993.00
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The AllPrep DNA/RNA FFPE Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Want to try this solution for the first time?
Get in touch with our team today and request a quote to trial the AllPrep DNA/RNA FFPE Kit (50).

Features

  • Maximum output with minimal sample consumption
  • Releases DNA/RNA without compromising integrity
  • Effectively separates RNA and DNA
  • Comprehensive DNA and RNA analysis of the same FFPE sample

Product Details

For reliable comparison of genomic and transcriptomic data, purification of DNA and RNA from the same sample is essential. The AllPrep DNA/RNA FFPE Kit uses a patent-pending solubilization method to purify DNA and RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. Purified analytes are suitable for use in applications such as real-time PCR and Pyrosequencing. The kit can be automated on the QIAcube Connect.

Performance

DNA and RNA purified using the AllPrep DNA/RNA FFPE Kit are of comparable quality to DNA and RNA purified using the QIAamp FFPE Tissue Kit and RNeasy FFPE Kit/miRNeasy FFPE, respectively (see figure " Purification of DNA and RNA from FFPE samples"). The purified nucleic acids are therefore suitable for downstream applications such as Pyrosequencing or real-time PCR and RT-PCR (see figure " Reliable amplification of DNA and RNA from FFPE samples" and " Array analysis following preamplification of cDNA targets").
See figures

Principle

The AllPrep DNA/RNA FFPE Kit is specially designed for simultaneous purification of genomic DNA and total RNA from FFPE tissue sections. Pure DNA and RNA are obtained from the entire sample, in contrast to other procedures where the biological sample is divided into two before being processed separately. Simply dividing a sample in half for separate DNA and RNA purification procedures results in the purification of DNA and RNA from different populations of cells, which may differ in their properties. Purification of DNA and RNA from the same sample also helps to prevent waste, since FFPE samples are precious, often difficult to retrieve, and limited in amount.

Due to fixation and embedding conditions, nucleic acids in FFPE samples are usually heavily fragmented and are often of a lower molecular weight than those obtained from fresh or frozen samples. A major obstacle to isolating DNA and RNA from the same FFPE sample is that fragmented DNA is short and can be partly single-stranded and therefore more closely resembles RNA than intact DNA. This property of fragmented DNA makes physical separation of DNA and RNA difficult. The AllPrep DNA/RNA FFPE Kit uses a patent-pending solubilization method to differentially release DNA and RNA from a single FFPE sample.

Nucleic acids in FFPE samples are also chemically modified by formaldehyde. Although formaldehyde modification cannot be detected in standard quality control assays, such as gel electrophoresis or lab-on-a-chip analysis, it does strongly interfere with enzymatic analyses. The AllPrep DNA/RNA FFPE Kit is optimized to reverse as much formaldehyde modification as possible without further DNA and RNA degradation.

Procedure

A simple workflow allows the purification of high-quality DNA and RNA from the same FFPE tissue section sample (see flowchart " AllPrep DNA/RNA FFPE procedure"). The AllPrep DNA/RNA FFPE Kit uses a patent-pending solubilization method to differentially release DNA and RNA from a single FFPE sample. With this method, FFPE samples are incubated in an optimized lysis buffer, which results in the release of RNA and precipitation of DNA. After centrifugation, the RNA-containing supernatant and DNA-containing pellet are then processed separately to purify RNA and DNA. Further incubations partially reverse crosslinking, and RNA or DNA is then purified using an RNeasy MinElute spin column or QIAamp MinElute spin column. For purified RNA, an on-column DNase treatment efficiently removes any contaminating DNA. Depending on the RNA binding conditions, small RNAs such as miRNA are either absent or present in the purified RNA. For purified DNA, an on-column RNase treatment is optional, as RNA contamination is minimal due to the separation of DNA and RNA prior to spin column processing.
See figures

Applications

The AllPrep DNA/RNA FFPE Kit is optimized to reverse as much formaldehyde modification as possible without further DNA and RNA degradation; however nucleic acids purified from FFPE samples should not be used in downstream applications that require high-molecular-weight DNA or full-length RNA. Some applications may require modifications to allow the use of fragmented nucleic acids (e.g., designing small amplicons for PCR and RT-PCR). For cDNA synthesis, gene-specific primers should be used instead of oligo-dT primers. If it is not possible to use gene-specific primers, random primers should be used.

Supporting data and figures

Specifications

FeaturesSpecifications
applicationsPCR, qPCR, real-time RT-PCR, microarray
elutionvolumeRNA: 14-30µl ; DNA: 30-100µl
purificationoftotalrnamirnapolyamrnadnaorproteinRNA (miRNA) and DNA
processingManual (centrifugation)
formatSpin column
numberofprepsperrun50
sampleamountmax. 4*10 µm sections or 2*20 µm sections
technologySilica technology
timeperrunorperprep6h for 10 samples, including sectioning
yieldVaries
mainsampletypeFFPE tissue samples

Resources

Brochures & Guides en (3)
Sample to Insight solutions for successful molecular analysis
High-quality, nucleic acid purification for successful PCR and NGS experiments.
Critical factors for molecular analysis of FFPE samples
Quick-Start Protocols (2)
Kit Handbooks (1)
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Additional Resources (1)
Certificates of Analysis (1)

Publications

Neoadjuvant atezolizumab for resectable non-small cell lung cancer: an open-label, single-arm phase II trial.
Chaft JE; Oezkan F; Kris MG; Bunn PA; Wistuba II; Kwiatkowski DJ; Owen DH; Tang Y; Johnson BE; Lee JM; Lozanski G; Pietrzak M; Seweryn M; Byun WY; Schulze K; Nicholas A; Johnson A; Grindheim J; Hilz S; Shames DS; Rivard C; Toloza E; Haura EB; McNamee CJ; Patterson GA; Waqar SN; Rusch VW; Carbone DP; LCMC study investigators;
Nat Med; 2022; 28 (10):2155-2161 2022 Sep 12 PMID:36097216
Personalized neoantigen vaccine NEO-PV-01 with chemotherapy and anti-PD-1 as first-line treatment for non-squamous non-small cell lung cancer.
Awad MM; Govindan R; Balogh KN; Spigel DR; Garon EB; Bushway ME; Poran A; Sheen JH; Kohler V; Esaulova E; Srouji J; Ramesh S; Vyasamneni R; Karki B; Sciuto TE; Sethi H; Dong JZ; Moles MA; Manson K; Rooney MS; Khondker ZS; DeMario M; Gaynor RB; Srinivasan L;
Cancer Cell; 2022; 40 (9):1010-1026.e11 2022 Aug 25 PMID:36027916
Transcriptome analysis reveals tumor microenvironment changes in glioblastoma.
Hoogstrate Y; Draaisma K; Ghisai SA; van Hijfte L; Barin N; de Heer I; Coppieters W; van den Bosch TPP; Bolleboom A; Gao Z; Vincent AJPE; Karim L; Deckers M; Taphoorn MJB; Kerkhof M; Weyerbrock A; Sanson M; Hoeben A; Lukacova S; Lombardi G; Leenstra S; Hanse M; Fleischeuer REM; Watts C; Angelopoulos N; Gorlia T; Golfinopoulos V; Bours V; van den Bent MJ; Robe PA; French PJ;
Cancer Cell; 2023; 41 (4):678-692.e7 2023 Mar 9 PMID:36898379
Detection of early seeding of Richter transformation in chronic lymphocytic leukemia.
Nadeu F; Royo R; Massoni-Badosa R; Playa-Albinyana H; Garcia-Torre B; Duran-Ferrer M; Dawson KJ; Kulis M; Diaz-Navarro A; Villamor N; Melero JL; Chapaprieta V; Dueso-Barroso A; Delgado J; Moia R; Ruiz-Gil S; Marchese D; Giró A; Verdaguer-Dot N; Romo M; Clot G; Rozman M; Frigola G; Rivas-Delgado A; Baumann T; Alcoceba M; González M; Climent F; Abrisqueta P; Castellví J; Bosch F; Aymerich M; Enjuanes A; Ruiz-Gaspà S; López-Guillermo A; Jares P; Beà S; Capella-Gutierrez S; Gelpí JL; López-Bigas N; Torrents D; Campbell PJ; Gut I; Rossi D; Gaidano G; Puente XS; Garcia-Roves PM; Colomer D; Heyn H; Maura F; Martín-Subero JI; Campo E;
Nat Med; 2022; 28 (8):1662-1671 2022 Aug 11 PMID:35953718
Multiomics in primary and metastatic breast tumors from the AURORA US network finds microenvironment and epigenetic drivers of metastasis.
Garcia-Recio S; Hinoue T; Wheeler GL; Kelly BJ; Garrido-Castro AC; Pascual T; De Cubas AA; Xia Y; Felsheim BM; McClure MB; Rajkovic A; Karaesmen E; Smith MA; Fan C; Ericsson PIG; Sanders ME; Creighton CJ; Bowen J; Leraas K; Burns RT; Coppens S; Wheless A; Rezk S; Garrett AL; Parker JS; Foy KK; Shen H; Park BH; Krop I; Anders C; Gastier-Foster J; Rimawi MF; Nanda R; Lin NU; Isaacs C; Marcom PK; Storniolo AM; Couch FJ; Chandran U; Davis M; Silverstein J; Ropelewski A; Liu MC; Hilsenbeck SG; Norton L; Richardson AL; Symmans WF; Wolff AC; Davidson NE; Carey LA; Lee AV; Balko JM; Hoadley KA; Laird PW; Mardis ER; King TA; AURORA US Network; Perou CM;
Nat Cancer; 2022; 4 (1):128-147 2022 Dec 30 PMID:36585450