Illumina HiSeq 2000 sequencing; Changes in the coding/non-coding transcriptome and DNA methylome that define the Schwann cell repair phenotype after nerve injury
Changes in the coding/non-coding transcriptome and DNA methylome that define the Schwann cell repair phenotype after nerve injury
Sciatic nerves of adult mice were cut or crushed at the sciatic notch. RNA was extracted from samples using Trizol (Invitrogen) according to manufacturer’s instructions with an additional purification step by on-column DNase treatment using the RNase-free DNase Kit (Qiagen) to ensure elimination of any genomic DNA. The integrity and quantity of RNA was determined using Qubit (Invitrogen) and Agilent 2100 Bioanalyzer (Agilent Technologies). Small RNA libraries were prepared using TruSeq Small RNA sample preparation kit (Illumina). Briefly, 3` and 5` indexed adapters were ligated to 1 g of total RNA and the product of this reaction was RT-PCR amplified. Small RNA library was size selected using Pippin Prep elution system (Sage Science). The quality of the libraries was evaluated using Agilent 2100 Bioanalyzer. All the libraries were pooled together and run on two lanes of the HiSeq 2000 platform to generate 50 bp single-end reads.
Experimental Factor: microRNA profiling by high throughput sequencing
protocol
Experimental Factor: cut nerve
injury
Experimental Factor: 3
time