Library preparation and sequencing were completed at ServiceXS, Plesmanlaan 1D, 2333 BZ, Leiden, Netherlands. The Illumina (San Diego, California, USA) mRNA-Seq and TruSeq Small RNA-Seq sample preparation kits were used to prepare sequencing libraries of mRNA and small RNA, respectively. Briefly, mRNA was selected by oligo-dT magnetic beads, fragmented and subjected to cDNA synthesis. Sequencing adapters were ligated to the cDNA fragments, followed by PCR amplification. Small RNA samples were processed by size exclusion gel electrophoresis subsequent to sequencing adaptor ligation. After gel excision and digestion, sequences were amplified by PCR. Clustering and DNA sequencing was performed using the Illumina cBot and HiSeq 2500. Each library was subjected to paired-end sequencing, producing reads 125 nucleotides in length.
Truseq small rna seq sample preparation kits
Manufactured by Illumina
Sourced in United States
The TruSeq Small RNA-Seq sample preparation kits are used to prepare small RNA samples for sequencing on Illumina platforms. The kits include reagents and protocols for the isolation, purification, and library preparation of small RNA molecules, such as microRNAs, from various sample types.
Automatically generated - may contain errors
Lab products found in correlation
Hiseq 2500, by Illumina (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Rna seq, by Illumina (1 mentions)
Qubit 2.0 fluorometer, by Thermo Fisher Scientific (1 mentions)
Qiazol lysis reagent, by Qiagen (1 mentions)
Mirneasy mini kit, by Qiagen (1 mentions)
2100 bioanalyser, by Agilent Technologies (1 mentions)
2 protocols using truseq small rna seq sample preparation kits
1
Illumina mRNA-Seq and Small RNA-Seq Library Preparation
2
Isolation and Sequencing of mRNA and Small RNA
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For RNA isolation frozen tissue or cultured cells were homogenized with Qiazol Lysis Reagent (Qiagen). The total RNA including the microRNA (miRNA) fraction was isolated using the miRNeasy Mini kit (Qiagen) according to manufacturer’s instructions. The concentration of RNA was determined using a NanoDrop™ 2000 spectrophotometer (Thermo Fisher Scientific) for cell cultures or Qubit® 2.0 Fluorometer (Life Technologies) for frozen tissue. For RNA-Seq the RNA integrity was assessed using a Bioanalyser 2100 (Agilent). Library preparation and sequencing were completed at GenomeScan. The Illumina RNA-Seq and TruSeq Small RNA-Seq sample preparation kits were used to prepare sequencing libraries of mRNA and small RNA in accordance to manufacturers guidelines. Clustering and DNA sequencing was performed using the Illumina cBot and HiSeq 2500 according to manufacturer’s protocols. Each library was subjected to paired-end sequencing, producing reads of 125 nucleotides in length with a read-depth of 36 million reads for RNA-Seq and 12 million reads for small RNA-Seq.
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