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  Prospects of transcript profiling for mRNAs and microRNAs using formalin-fixed and paraffin-embedded dissected autoptic multiple sclerosis lesions

Eisele, S., Krumbholz, M., Fischer, M.-T., Mohan, H., Junker, A., Arzberger, T., et al. (2012). Prospects of transcript profiling for mRNAs and microRNAs using formalin-fixed and paraffin-embedded dissected autoptic multiple sclerosis lesions. Brain Pathology, 22(5), 607-618. doi:10.1111/j.1750-3639.2012.00564.x.

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 Creators:
Eisele, Sylvia1, Author           
Krumbholz, Markus1, Author           
Fischer, Marie-Therese, Author
Mohan, Hema1, Author           
Junker, Andreas1, Author           
Arzberger, Thomas, Author
Hohlfeld, Reinhard1, Author           
Bradl, Monika, Author           
Lassmann, Hans, Author
Meinl, Edgar1, Author           
Affiliations:
1Department: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society, ou_1113547              

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Free keywords: POLYMERASE-CHAIN-REACTION; IN-SITU HYBRIDIZATION; BARR-VIRUS INFECTION; GENE-EXPRESSION; UP-REGULATION; HUMAN-BRAIN; RT-PCR; TISSUES; POSTMORTEM; STABILITYFFPE; microRNA; molecular analysis; multiple sclerosis; quantitative PCR; transcriptome;
 Abstract: The elaboration of novel pathogenic aspects of multiple sclerosis (MS) requires the analysis of well-defined stages of lesion development. However, specimens of certain stages and lesion types are either present in small brain biopsies, insufficient in size for further molecular studies or available as formalin-fixed and paraffin-embedded (FFPE) material only. Therefore, application of current molecular biology techniques to FFPE tissue is warranted. We compared FFPE and frozen tissue by using quantitative polymerase chain reaction and report: (1) FFPE material is highly heterogeneous regarding the utility for transcript profiling of mRNAs; well-preserved FFPE samples had about a 100-fold reduced sensitivity compared with frozen tissue, but gave similar results for genes of sufficient abundance; (2) FFPE samples not suitable for mRNA analysis are still highly valuable for miRNA quantification; (3) the length of tissue fixation greatly affects utility for mRNA but not for miRNA analysis; (4) FFPE samples can be processed via a hot water bath for dissection of defined lesion areas; and (5) in situ hybridization for proteolipid protein (PLP) helps to identify samples not suitable for mRNA amplification. In summary, we present a detailed protocol how to use autoptic FFPE tissue for transcript profiling in dissected tissue areas.

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Language(s): eng - English
 Dates: 2012-09
 Publication Status: Published in print
 Pages: 12
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

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Title: Brain Pathology
  Other : Brain Pathol.
Source Genre: Journal
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Publ. Info: Zürich, Switzerland : International Society of Neuropathology
Pages: - Volume / Issue: 22 (5) Sequence Number: - Start / End Page: 607 - 618 Identifier: ISSN: 1015-6305
CoNE: https://pure.mpg.de/cone/journals/resource/954925585260