The Microarray and Genomic Analysis Core Facility provides access to an Illumina Genome Analyzer, which enables massively parallel DNA sequencing.
The resource is operated as a full service facility that provides experimental handling of all aspects of the Illumina sequencing processes.
HiSeq2000 101 Cycle Paired End Sequencing - $2925.00 per lane
HiSeq2000 50 Cycle Single Read Sequencing - $1260.00 per lane
HiSeq2000 50 Cycle Paired End Sequencing - $2115.00 per lane
All genomic DNA samples submitted to the core facility for library construction will go through quality control (NanoDrop spectrophotometer reading and evaluate quality on a 1% agarose gel) prior to library construction.
Barcode: All libraries constructed at the core facility will include an index barcode tag which enables multiplex sequencing if desired. qPCR is performed on all samples prior to combining indexed samples that will be run in the same lane to provide a more accurate concentration of cluster forming units in the library. Although the intention of qPCR is to equalize the molarity of all samples that contribute to a single lane, it is not perfect and one should expect that at times there will be up to a 20% deviation in the sample that is most highly represented and the one that is most lowly represented.
Genomic DNA libraries are size selected with an average insert size of 300-400 bp. If you would like a smaller or larger size range, please let us know.
Post-Library Quality Control includes the following services: (1)NanoDrop reading, (2)Q-pcr quantitation of library concentration using Illumina primers, and (3) evaluation of library on a Agilent Bioanalyzer DNA 1000 chip. These services are included when the library is constructed at the core facility. If the library has already been constructed when submitted to the core facility, a post-library quality control charge will be added to the charges for the sequencing.
Most genomic DNA libraries are sequenced on a 101 cycle Paired End sequencing run. However, the client can select whatever run type is needed for their project.
Version 3 flowcells used for paired end sequencing currently have a specification of delivering 240 to 300 GB of passed filter reads per flowcell on a 101 cycle paired end run. There are 8 lanes on each flowcell and therefore, Illumina's specification per lane of 101 cycle paired end sequencing is 30-37.5 GB of passed filter data per lane. Illumina's specification is based on samples that have previously been run at least one time since this allows you to optimize the cluster number on future runs. We attempt to meet this specification on the first run for each lane of samples.
Version 1.5 flowcells are currently used for single end read sequencing and these have a specification of delivering 18.75 to 25 GB of passed filter reads per flowcell on a 50 cycle single end read run. There are 8 lanes on each flowcell and therefore Illumina's specification per lane of 50 cycle single end sequencing is 2.3 to 3.1 GB of passed filter data per lane. Illumina's specification is based on samples that have previously been run at least one time since this allows you to optimize the cluster number on future runs. We attempt to meet this specification on the first run for each lane of samples.
Illumina sequencing flowcells contain eight lanes and all eight lanes must be filled prior to starting the sequence run. We seldom receive sequencing requests for 101 cycle single read or for 50 cycle paired end sequencing.
Therefore, the waiting period for sequence analysis on one of these two run types can be lengthy. It is advised to contact the core facility prior to requesting such a run type or to plan on filling out all eight lanes of the flowcell so that the run can start in a timely manner.
A PhiX control library that is constructed by Illumina is spiked into each lane at a concentration that represents approximately 0.5% of the reads.
Genomic DNA Library Prep
Microarray and Genomic Analysis Core Facility has not received any reviews.
Microarray and Genomic Analysis Core Facility has not received any endorsements.