ScriptSeq Complete Kit (Human/Mouse/Rat)

RNA sequencing Mouse - J774

Experiment
RNA sequencing Mouse - J774
Product
ScriptSeq Complete Kit (Human/Mouse/Rat) from Illumina
Manufacturer
Illumina

Protocol tips

Protocol tips
The Ribo-Zero process uses 100 ng - 1 μg of DNA-free RNA.

Publication protocol

Approximately 500 ng of RNA from each sample was used to prepare individually barcoded strand-specific RNA-seq libraries. rRNA and mitochondrial RNA was removed using Ribo-Zero Gold Removal Magnetic Kit for human/mouse/rat (Epicentre, now Illumina) following the manufacturer’s instructions. mRNA was purified using Agencourt RNAClean XP kit. Libraries were constructed using ScriptSeq™ Complete Kit (Epicentre, now Illumina) as per manufacturer’s instructions. Quality and quantity of the libraries were determined using a Bioanalyzer (Agilent) and Qubit® (Invitrogen), respectively. Libraries were then quantified by qPCR in an Applied Biosystems® 7500 with the Kapa Biosystems kit for library quantitation (Kapa Biosystems). Next-generation sequencing was performed using an Illumina HiSeq 2000 flow cell with one 150-bp end run. The NCBI Gene Expression Omnibus (GEO) accession number for the RNA-seq data reported in this paper is GSE88801.

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Discussion

Discussion

5 years ago

Author: Panagiotis Karagiannis Greece

Using RNA from formalin fixed paraffin embedded tissue sections for sequencing

Hello there! I have some slides with mouse lung tumors and I would like to do rna sequencing. Will I be able to extract enough/good quality rna from RNA from formalin fixed paraffin embedded tissue sections for sequencing to be successful?

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Papers

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Manufacturer protocol

Download the product protocol from Illumina for ScriptSeq Complete Kit (Human/Mouse/Rat) below.

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