Illumina MiSeq Desktop Sequencer
The Translational Genomics Core at Partners HealthCare Personalized Medicine features the Illumina MiSeq Desktop Sequencer. The MiSeq offers a faster turn-around time, with the ability to complete sequencing in a matter of hours. It delivers the same high-quality data with Illumina’s sequencing-by-synthesis technology. This sequencer is best suited for whole genome sequencing of smaller model organism genomes, medium to small targeted panels, amplicon sequencing, and for quality control of libraries. We guarantee a minimum of 10 million clusters per lane of raw, unfiltered data on the MiSeq regular flowcell.
Researchers can submit pre-made libraries or purified DNA/RNA for library construction and subsequent sequencing. The Translational Genomics Core will also perform quality control (QC) on all libraries by qPCR prior to sequencing.
Pooling of libraries is available at an additional cost. Pooling information should be added under special service instructions when submitting a GIGPAD batch order into our online LIMS.
Sample Requirements for Customer-Constructed Libraries
|Sample Type||Minimum Volume||Solution||Concentration||Fragment Size||Purification|
|Customer-constructed libraries||20 ul||TE or nuclease free water||20-100 nM (quantiated by picogreen)||Electrophoresis Gel picture of BioAnalyzer profile should be included||Qiagen QIAquick PCR purification (28104) or Agencourt Ampure XP (A63881) kit|
Please submit samples in either VWR free-standing RNase/DNase free 0.5ml tubes (89004-286) or Fisher RNase/DNase free plates (AB-0800). We can provide these tubes/plates free of charge. If samples are submitted in a different container type, there will be a $5 transfer charge per sample.
All samples submitted will be ensured for quality control (QC) to check concentration and purity by optical density (OD) using a Nanodrop 1000 and Picogreen assay using a Qubit Fluorometer. In the case of total RNA, RIN will be analyzed using Agilent Bioanalyzer chips or Tapestation tape. Any sample failing QC will be returned to the customer for replacement. Where no replacement is available, we can process the failing samples but cannot guarantee the success of the library construction, and no attempt to troubleshoot can be made.
We work on a first-come, first-served basis and queue libraries only after passing QC. Faster turn-around time can be offered to customers who fill full flow cells on the high-capacity mode or opt to sequence in the rapid-run mode. Once on the flowcell, the rapid-run mode offers the fastest sequencing time of 24–48 hours. Depending on the read length, the high-capacity mode takes 5–12 days to sequence once on the flowcell.