Library Preparations

We provide both DNA and RNA library preparation services for a wide variety of Next-Generation Sequencing applications. Consult with our team to determine which approach is right for you.

Consultations

We use Illumina-compatible library preparation kits to ensure quality data from sequencing runs. Below are some of the common applications and corresponding prep kits we regularly have in stock.

Coming soon: 10x Genomics Chromium Next GEM single cell 3' library preparation

Contact CHGI for more information and to start planning your single cell RNAseq projects.

Don't see your required library type listed below?

Book a consultation with us. We do our best to take custom requests.

Sample submission tips

To get an idea of the required sample submission concentration and volume, please see the information found on the "Sample Requirements" tab in the Sample Submission Form. Make sure to follow these general guidelines:

Ensure your nucleic acid is free from contaminants;
- DNA samples may need to be treated with RNase, and RNA samples with DNase
Samples should be suspended in nuclease-free water or 10 mM Tris only;
- do not suspend samples in buffer containing EDTA, it will inhibit downstream reactions.
Submit samples in 200-μL strip tubes or 96-well plates.

Whole Genome Sequencing

Genomic DNA is sheared by Covaris sonicators or enzymatically fragmented to prepare DNA fragment libraries. More input material is preferable to reduce the number PCR cycles required for amplification. Alternatively, PCR-free workflows are available for human WGS or shotgun metagenomics.

Popular WGS Library Prep Workflows:

Whole Exome Sequencing and Custom Capture Targets

The protein-coding region is captured using biotinylated RNA or DNA baits to enrich for all exons. Optional untranslated regions (UTRs) or catalog of somatic mutations in cancer (COSMIC) regions can also be included. We commonly stock IDT's xGen Exome Research Panel v2 and Agilent’s Clinican Research Exome v2 for whole human exome sequencing, but non-human exome sequencing for mouse, bovine, and zebrafish can be ordered. Alternatively, Arbor Biosciences provides MyBait RNA probes which can be custom-designed to target regions in the genomes of a wide range of organisms. These are ordered and supplied by the customer on a per-project basis.

Popular Exome and Custom Capture Workflows:

IDT xGen Exome Research Panel v2
Agilent SureSelect XT2 Clinical Research Exome v2
MyBaits custom target capture, user-supplied capture probes

16S Metagenomics and Amplicon Sequencing

The V3-V4 region of microbial 16S rRNA genes is amplified by PCR, and barcoded in a second round of PCR with Nextera XT2 dual indexes. For bacterial/fungal population mixes, a custom 16S/ITS panel can be used for community profiling. Other regions of interest can be amplified, indexed and sequenced using custom-designed primers.

Library Prep Protocols

Shotgun Metagenomics

As an alternative to sequencing part of the 16S rRNA genes, an entire population of microbial genomic DNA can be sequenced as a single shotgun DNA fragment library.

Popular Shotgun Metagenomics Library Prep Workflows:

ChIP-Seq

After immunoprecipitation, ChIP-Seq DNA fragments are ~100-300 bp, so shearing isn’t required. Fragment libraries are constructed from as little as a few nanograms of input material.

Library Prep Protocol:

NEBNext Ultra II DNA Library Prep Kit

Circulating Cell-Free DNA or FFPE DNA

Circulating nucleic acids (CNA) are size-selected by magnetic bead clean-up, and converted to fragment libraries. Selected regions can be enriched by PCR amplification. FFPE DNA may also be used as input into the IDT xGen cfDNA & FFPE DNA Library Prep workflow.

Popular Library Prep Protocols:

Stranded poly(A) RNA-Seq

Our most popular RNA-seq service isolates mRNA molecules for gene expression profiling and discovery.

Popular Library Prep Protocols:

NEBNext Ultra II Directional RNA Library Prep Kit with poly(A) selection
Illumina Stranded mRNA Prep Ligation

Stranded Total RNA-Seq

We can also prepare total RNA-seq libraries with rRNA depletion for whole transcriptome analysis. We offer options for ultra-low input and degraded RNA samples (typically extracted from FFPE tissue).