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Next-Gen Sequencing

Applications of Next-Gen Sequencing

Next-generation (massively parallel) sequencing is useful for a variety of experimental approaches. It is not a replacement for normal (Sanger) sequencing of plasmids or PCR products. Instead, next-gen sequencing relies on capturing millions or billions of individual DNA molecules (e.g. genomic DNA fragments, cDNAs), which are then amplified separately and sequenced in parallel, generating millions or billions of sequencing "reads", each of which originated from a different template molecule. It is similar to individually cloning and sequencing millions or billions of independent DNA fragments, but it all happens at once and in just a few days.

The following types of scientific applications are easily adaptable to next-gen sequencing approaches:

• Targeted gene panel sequencing of cancer- or disease-relevant genes
• Gene expression studies using RNA-seq
• Chromatin Immunoprecipitation - sequencing (ChIP-seq) for transcription factor or epigenetic studies
• DNA methylation studies (e.g. RRBS)
• Transcriptome sequencing (e.g. identification of alternatively-spiced RNAs)
• Analysis of non-coding RNAs (ncRNAs, lincRNAs, miRs)

Technologies and Instruments

The ATG facility currently has or has access to several types of next-gen sequencing instruments:

• Ion S5/XL: A solid state NGS insturment generating up to 100 million reads per chip

These instruments provide cost-efficient and rapid next-generation sequencing for applications such as targeted sequencing, RNA-seq, ChIP-seq, MeDIP-seq and even low-pass whole-genome sequencing for copy number variant analysis.

Please contact the ATG Facility staff for more information about assays and pricing.