SGT RNA Sequencing Services

mRNA-sequencing (mRNA-seq) is a popular tool for quantifying gene expression levels from polyadenylated mRNA transcripts. mRNA-Seq can also interrogate alternatively spliced variants to help identify novel isoforms. We offer complete solutions for mRNA-Seq projects including experimental design, RNA-seq library preparation, high throughput Illumina sequencing, and data processing.

Transcriptomes can also be assembled without the need of a reference genome. This approach is useful for determining gene sequences of non-model organisms. We offer services for performing whole transcriptome de novo sequencing on Illumina platforms.

The correct kit, read length, and depth will depend on the needs of your project. Please contact us for help with experimental design.  

Total RNA-Seq with ribosomal reduction selectively removes ribosomal RNA from total RNA samples. In contrast to polyA⁺ enrichment mRNA-seq, this approach preserves non-polyadenylated RNAs to investigate broader classes of RNAs including immature mRNAs and non-polyadenylated ncRNAs. For most samples, we prepare RNA-seq libraires using the Illumina TruSeq Stranded total RNA-seq kit in combination with a wide variety of ribosomal reduction kits, which removes cytoplasmic and mitochondrial rRNAs from human/mouse/rat/bacterial samples. Custom rRNA reduction kits can be ordered for yeast and other organisms.

Small/mi RNA analysis provides the ability to discover, measure and compare expression of know microRNAs and other small non-coding RNA. It also allows detection of novel microRNA targets. We use the Qiagen miRNA Library Prep kit  for all miRNA library preps. 

Ultra-Low-Input RNA-Seq can be used to generate gene expression data from few or even single cells. Ultra-low-input RNA-Seq provides the opportunity to study differences between cells or cell types with an unprecedented resolution. We use the Clontech Ultra Low Input RNA SMARTer mRNA amplification kit for the initial RNA amplification, which provides a fast and simple method for preparing amplified cDNA from total RNA for RNA-Seq applications. We convert this double stranded cDNA into an Illumina sequencing library using Kapa Hyper Prep Kit.

Studying gene expression of FFPE samples is extremely valuable for better understanding diseases. Unfortunately, FFPE samples can be significantly degraded, and traditional RNA-seq library preparation methods often fail to generate optimal libraries. Depending on the degradation of the RNA, we can use the Illumina total RNA kit or RNA Exome kit, an alternative sample preparation method enabling the generation of high quality RNA-Seq results from degraded samples.