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Laser capture microdissection for transcriptomic profiles in human skin biopsies

Overview of attention for article published in BMC Molecular Biology, June 2018
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Title
Laser capture microdissection for transcriptomic profiles in human skin biopsies
Published in
BMC Molecular Biology, June 2018
DOI 10.1186/s12867-018-0108-5
Pubmed ID
Authors

Silvia Santoro, Ignazio Diego Lopez, Raffaella Lombardi, Andrea Zauli, Ana Maria Osiceanu, Melissa Sorosina, Ferdinando Clarelli, Silvia Peroni, Daniele Cazzato, Margherita Marchi, Angelo Quattrini, Giancarlo Comi, Raffaele Adolfo Calogero, Giuseppe Lauria, Filippo Martinelli Boneschi

Abstract

The acquisition of reliable tissue-specific RNA sequencing data from human skin biopsy represents a major advance in research. However, the complexity of the process of isolation of specific layers from fresh-frozen human specimen by laser capture microdissection, the abundant presence of skin nucleases and RNA instability remain relevant methodological challenges. We developed and optimized a protocol to extract RNA from layers of human skin biopsies and to provide satisfactory quality and amount of mRNA sequencing data. The protocol includes steps of collection, embedding, freezing, histological coloration and relative optimization to preserve RNA extracted from specific components of fresh-frozen human skin biopsy of 14 subjects. Optimization of the protocol includes a preservation step in RNALater® Solution, the control of specimen temperature, the use of RNase Inhibitors and the time reduction of the staining procedure. The quality of extracted RNA was measured using the percentage of fragments longer than 200 nucleotides (DV200), a more suitable measurement for successful library preparation than the RNA Integrity Number (RIN). RNA was then enriched using the TruSeq® RNA Access Library Prep Kit (Illumina®) and sequenced on HiSeq® 2500 platform (Illumina®). Quality control on RNA sequencing data was adequate to get reliable data for downstream analysis. The described implemented and optimized protocol can be used for generating transcriptomics data on skin tissues, and it is potentially applicable to other tissues. It can be extended to multicenter studies, due to the introduction of an initial step of preservation of the specimen that allowed the shipment of biological samples.

Twitter Demographics

The data shown below were collected from the profile of 1 tweeter who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 27 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 27 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 6 22%
Researcher 5 19%
Other 2 7%
Student > Bachelor 2 7%
Student > Postgraduate 2 7%
Other 2 7%
Unknown 8 30%
Readers by discipline Count As %
Neuroscience 5 19%
Medicine and Dentistry 5 19%
Biochemistry, Genetics and Molecular Biology 4 15%
Social Sciences 2 7%
Agricultural and Biological Sciences 1 4%
Other 3 11%
Unknown 7 26%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 20 June 2018.
All research outputs
#11,645,444
of 13,110,606 outputs
Outputs from BMC Molecular Biology
#218
of 263 outputs
Outputs of similar age
#232,143
of 268,114 outputs
Outputs of similar age from BMC Molecular Biology
#1
of 1 outputs
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