Comparison of Two Methods of RNA Extraction from Formalin-Fixed Paraffin-Embedded Tissue Specimens

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dc.contributor Sistema FMUSP-HC: Faculdade de Medicina da Universidade de São Paulo (FMUSP) e Hospital das Clínicas da FMUSP GOUVEIA, Gisele Rodrigues FMUSP-HC
FERREIRA, Jerenice Esdras FMUSP-HC
SIQUEIRA, Sheila Aparecida Coelho FMUSP-HC
PEREIRA, Juliana FMUSP-HC 2014
dc.identifier.citation BIOMED RESEARCH INTERNATIONAL, article ID 151724, 5p, 2014
dc.identifier.issn 2314-6133
dc.description.abstract The present study aimed to compare two different methods of extracting RNA from formalin-fixed paraffin-embedded (FFPE) specimens of diffuse large B-cell lymphoma (DLBCL). We further aimed to identify possible influences of variables-such as tissue size, duration of paraffin block storage, fixative type, primers used for cDNA synthesis, and endogenous genes tested-on the success of amplification from the samples. Both tested protocols used the same commercial kit for RNA extraction (the Recover All Total Nucleic Acid Isolation Optimized for FFPE Samples from Ambion). However, the second protocol included an additional step of washing with saline buffer just after sample rehydration. Following each protocol, we compared the RNA amount and purity and the amplification success as evaluated by standard PCR and real-time PCR. The results revealed that the extra washing step added to the RNA extraction process resulted in significantly improved RNA quantity and quality and improved success of amplification from paraffin-embedded specimens.
dc.description.sponsorship · Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (State of Sao Paulo Research Support Foundation - FAPESP)
dc.language.iso eng
dc.relation.ispartof Biomed Research International
dc.rights openAccess
dc.subject.other dna; pcr; expression
dc.title Comparison of Two Methods of RNA Extraction from Formalin-Fixed Paraffin-Embedded Tissue Specimens
dc.type article
dc.rights.holder Copyright HINDAWI PUBLISHING CORPORATION LIM/46 LIM/14 LIM/21 LIM/31
dc.identifier.doi 10.1155/2014/151724
dc.identifier.pmid 25105117
dc.type.category original article
dc.type.version publishedVersion GOUVEIA, Gisele Rodrigues:FM:MPS FERREIRA, Suzete Cleusa:HC:LIM/46 FERREIRA, Jerenice Esdras:IMT: SIQUEIRA, Sheila Aparecida Coelho:HC:LIM/14 PEREIRA, Juliana:FM:MCM 2-s2.0-84904678547 WOS:000338852600001 NEW YORK USA
hcfmusp.relation.reference · Antica M, 2010, J IMMUNOL METHODS, V359, P42, DOI 10.1016/j.jim.2010.05.010
· Bonin S, 2005, J CLIN PATHOL, V58, P313, DOI 10.1136/jcp.2004.016477
· Doleshal M, 2008, J MOL DIAGN, V10, P203, DOI 10.2353/jmoldx.2008.070153
· Gillio-Tos A, 2007, PATHOLOGY, V39, P345, DOI 10.1080/00313020701329757
· Gouveia G. R., 2011, Jornal Brasileiro de Patologia e Medicina Laboratorial, V47, P649, DOI 10.1590/S1676-24442011000600012
· Hamatani K, 2006, J HISTOCHEM CYTOCHEM, V54, P773, DOI 10.1369/jhc.5A6859.2006
· Korbler T, 2003, EXP MOL PATHOL, V74, P336, DOI 10.1016/S0014-4800(03)00024-8
· Lehmann U, 2001, METHODS, V25, P409, DOI 10.1006/meth.2001.1263
· Li JH, 2008, BMC BIOTECHNOL, V8, DOI 10.1186/1472-6750-8-10
· Ribeiro-Silva A., 2008, Jornal Brasileiro de Patologia e Medicina Laboratorial, V44, P123, DOI 10.1590/S1676-24442008000200009
· Scorsato A. P., 2011, Jornal Brasileiro de Patologia e Medicina Laboratorial, V47, P541, DOI 10.1590/S1676-24442011000500008
· Thermo Scientific, 2008, NANODROP 1000 SPECTR
· Witchell J, 2008, PATHOL RES PRACT, V204, P105, DOI 10.1016/j.prp.2007.09.002
dc.description.index MEDLINE
dc.identifier.eissn 2314-6141
hcfmusp.citation.scopus 8
hcfmusp.citation.wos 8

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