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Next-generation sequencing - UNISI
Free
Services do NOT include
Costs for shipment of samples, additional Bioinformatic analyses.
Possible Output
Data will be provided in form of counts table (csv file) and/or bam files.
Sample Requirements - input of users
Frozen total RNA from any human sample and/or human derived cell-cultures. Minimum input RNA: 200 ng, minimum RNA concentration 10 ng/ul (quantified by a fluorimetric method, e.g. Qubit). RNA should preferably be suspended in water or 5mM Tris (pH 7.5-8.5) and shipped on dry ice in properly labelled DNAse-, RNAse-free 1.5 ml tubes. Users may also provide frozen PaxGene tubes containing blood from patients/volunteers.
Lead Scientist
Donata Medaglini
Description of service
UNISI will provide a targeted RNA sequencing service. If necessary, RNA will be extracted from PaxGene tubes. RNA will be quantified by fluorimetry, retrotranscribed and then sequenced using Ion AmpliSeq™ Transcriptome Human Gene Expression Kit on the “Ion Proton™” (Thermo Fisher) sequencing platform. The expression level of 20,802 human genes will be quantified. A minimum of 8 million reads/sample will be guaranteed. RNA obtained from any human sample can be sequenced with this protocol.
Timeline
2 weeks for every 16 RNA samples,
plus an 1 more week for every other 16 samples. Consider 1 additional week if providing PaxGene tubes.
Publications
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Li W, Turner A, Aggarwal P, Matter A, Storvick E, Arnett DK, Broeckel U. Comprehensive evaluation of AmpliSeq transcriptome, a novel targeted whole transcriptome RNA sequencing methodology for global gene expression analysis. BMC Genomics. 2015 Dec 16;16:1069. doi: 10.1186/s12864-015-2270-1. PubMed PMID: 26673413.
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