Next-generation sequencing - UNISI
Services do NOT include
Costs for shipment of samples, additional Bioinformatic analyses.
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.
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.
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.
The Department of Medical Biotechnology, Laboratory Molecular Microbiology and Biotechnology, University of Siena, Italy
0039 (0)577 233101
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.