Illumina Library Prep
GTAC@MGI generates libraries with many methods to fit your needs for either RNA-seq or DNA-seq.
RNA-Seq
What are my options for prep type?
Consider GTAC@MGI to facilitate your evaluation with RNA-Seq. This process will allow you to tell which genes are turned on, their level of expression, and the timing of activation. The first steps for the prep are dependent of your RNA quality, quantity, and experimental goals. Depending on your focus, we have a set of quality choices to choose from.
- PolyA selection
- Ribosomal depletion, Kapa RiboErase (H/M/R)
- Ribosomal depletion, Qiagen FastSelect (H/M/R)
- Ribosomal and globin depletion, Qiagen FastSelect (H/M/R+Globin)
- Ribosomal and globin depletion, Watchmaker (H/M/R+Globin)
- Low input – Takara-Clontech SMARTseq mRNA
- Low input – Sigma Seqplex
Other options and kits may be available, including the ability to submit cDNA or mRNA. For single cell services, please see here. We regularly work with total RNAs derived from human, mouse, and rat, but we can also work with RNA studies involving model organisms, plants, bacteria, viruses and others. Please inquire with us.
Our RNA-seq laboratory provides RNA and process QC measurements utilizing Agilent Bioanalyzer and/or Tapestation and Qubit.
What are my options for sequencing?
GTAC@MGI creates libraries that are uniquely dual-indexed with 10bp fragments. Doing so allows your sample libraries to be multiplexed onto the appropriate amount of Illumina NovaSeq X Plus 2×150 runs. While 30 million clusters are a typical target for most mammalian gene expression profiling studies, we can tailor the sequencing depth to fit the needs of your study. If you desire a dedicated run on a full flow cell, other read lengths, or different instrument, please reach out to discuss options.
Data analysis?
For no extra fees the GTAC@MGI will provide its standard RNA-seq analytical pipeline. This comprehensive analysis of RNA-seq data includes multiple levels of quality control, differential expression, and pathway analysis. Additional details can be found here.
DNA-Seq
What are my options for prep type?
Consider GTAC@MGI to facilitate your evaluation with DNA-Seq. This process will allow you to analyze the entire genomic sequence of an organism that has been previously classified, analyze the protein-coding regions of genes in the genome, or target a panel of genes of interest. Some assays also use unique molecular identifiers to validate rare variants. Depending on your focus, we have a set of quality choices to choose from.
- Whole Genome – KAPA Hyper PCR-Free
- Whole Genome – KAPA Hyper amplified
- Whole Genome – IDT Methyl-seq
- Whole Genome – Low input IDT cfDNA/FFPE kit
- Whole Exome – KAPA Hyper amplified with IDT Exome
- Custom Capture – KAPA Hyper amplified with custom panel (reach out regarding compatible panel designs)
We also support ChIPseq to analyze protein interactions by combining chromatin IP with sequencing to identify DNA binding sites. Another option is Amplicon-Seq for ultra-deep sequencing of PCR products for known regions of interest. One common amplicon assay we support in our PCR lab is Bacterial 16S sequencing for identification, classification, and quantitation of bacterial microbes at a species level. Please inquire with us for additional options.
What are my options for sequencing?
GTAC@MGI creates libraries that are uniquely dual-indexed with 10bp fragments. Doing so allows your sample libraries to be multiplexed onto the appropriate amount of Illumina NovaSeq X Plus 2×150 runs. We can tailor the sequencing depth to fit the needs of your study. If you desire a dedicated run on a full flow cell, other read lengths, or different instrument, please reach out to discuss options.
Data analysis?
For no extra fees the GTAC@MGI will provide its standard analytical pipeline for genome, exome, 16S, and chip data. This comprehensive analysis of DNA-seq data includes FASTQ files, BAM files, and variant/peak calls. Additional details can be found here.