Next Generation Sequencing (NGS)
NGS technology utilizes a massively parallel approach to quantitatively assess copy number and sequence changes in DNA or RNA samples. It has become the preferred method for integrated approaches that simultaneously measure transcript copy numbers and sequence- based alterations. Two technologies are currently available in the core that will best fit your project, namely Illumina NextSeq™ 500 and MiSeq Sequencing System.
Each instrument supports sequencing kits that offer a range of possible cycle numbers, for instance a 150 cycle kit or 300 cycle kit. The number of cycles in the kit indicates how many nucleotides can be incorporated in the growing chain during sequencing. A 150 cycle kit, for instance, can generate a 150 nt-long single end read, or 2×75 bp-long paired end reads. Use the 300 cycles kit if you require pair-end reads that are 150 bp long.
NextSeq kits come in two output: High Output generates around 400 Million reads per run and Mid Output generates around 130 Million reads per run. Please keep this in mind as you are making your kit selection in iLab. Our staff can assist you in selecting the appropriate sequencing kit for your study.
We encourage the use of Run QC to check the number of targeted single cell before proceeding to optimize the NextSeq RNA-seq run.