https://observatorio.fm.usp.br/handle/OPI/7376
Title: | Comparison of Two Methods of RNA Extraction from Formalin-Fixed Paraffin-Embedded Tissue Specimens |
Authors: | GOUVEIA, Gisele Rodrigues; FERREIRA, Suzete Cleusa; FERREIRA, Jerenice Esdras; SIQUEIRA, Sheila Aparecida Coelho; PEREIRA, Juliana |
Citation: | BIOMED RESEARCH INTERNATIONAL, article ID 151724, 5p, 2014 |
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. |
Appears in Collections: | Artigos e Materiais de Revistas Científicas - FM/MCM Artigos e Materiais de Revistas Científicas - FM/MPS Artigos e Materiais de Revistas Científicas - HC/ICHC Artigos e Materiais de Revistas Científicas - IMT Artigos e Materiais de Revistas Científicas - LIM/14 Artigos e Materiais de Revistas Científicas - LIM/21 Artigos e Materiais de Revistas Científicas - LIM/31 Artigos e Materiais de Revistas Científicas - LIM/46 Artigos e Materiais de Revistas Científicas - ODS/03 |
File | Description | Size | Format | |
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art_GOUVEIA_Comparison_of_Two_Methods_of_RNA_Extraction_from_2014.PDF | publishedVersion (English) | 1.9 MB | Adobe PDF | View/Open |
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