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Efficient cellular fractionation of RNA

ABSTRACT: The aim of this experiment was to develop new methods to extract pure fractions of nuclear and cytoplasmic RNA. We compared the results of nuclear and cytoplasmic RNA-sequencing to results of the total and polyA+ RNA-sequencing. Cytoplasmic, nuclear, total and polyA+ RNA was extracted from two human brain samples and sequenced on the SOLiD system. We analyzed the data to look for differences in levels of nascent transcription and mature mRNAs between the different samples.

INSTRUMENT(S): AB 5500xl Genetic Analyzer

ORGANISM(S): Homo sapiens

SUBMITTER: Adam Ameur

PROVIDER: E-MTAB-1898 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Efficient cellular fractionation improves RNA sequencing analysis of mature and nascent transcripts from human tissues.

Zaghlool Ammar A Ameur Adam A Nyberg Linnea L Halvardson Jonatan J Grabherr Manfred M Cavelier Lucia L Feuk Lars L

BMC biotechnology 20131113

<h4>Background</h4>The starting material for RNA sequencing (RNA-seq) studies is usually total RNA or polyA+ RNA. Both forms of RNA represent heterogeneous pools of RNA molecules at different levels of maturation and processing. Such heterogeneity, in addition to the biases associated with polyA+ purification steps, may influence the analysis, sensitivity and the interpretation of RNA-seq data. We hypothesize that subcellular fractions of RNA may provide a more accurate picture of gene expressio ...[more]

PMID: 24225116

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