At the GRCF High Throughput Sequencing Center, our goal is to provide the research community at Johns Hopkins University with access to ‘next generation’ sequencing platforms. We currently feature three HiSeq 2500 instruments and two MiSeq instruments, and an Ion Proton (currently in R&D). We do our best to sequence samples in a timely manner, but our first priority is high quality data.
We offer simple, per lane pricing, along with end to end service for whole exome/custom targeted projects, RNASeq, and whole genome.
Per Lane Pricing
50 cycles:$1,200 per lane
100 cycles:$1,600 per lane
150 cycles:$2,000 per lane
Paired End Reads:
50 cycles (x2): $1,900 per lane
100 cycles (x2): $2,400 per lane
150 cycles (x2): $2,900 per lane
Non-standard read lengths are also available, provided they are submitted in pairs (enough to start a 2500 run). Please inquire about pricing.
Library Prep: $250/sample
Completed Libraries: Please submit 10μl of your sample at 2nM. Samples must be pooled. We will do a QC check via Bioanalyzer to increase the likelihood of quality data, but you should quantitate your sample as accurately as possible. Nanodrop is not reliable. qPCR is by far the most accurate, but intercalating dye methods can be used.
Library Prep: Please provide us with >500ng of high molecular weight DNA/RNA. Please contact us to discuss sample submission.
Our facility features the HiSeq 2500 platform. We almost exclusively run our instruments in Rapid Run Mode, due to the fast run times and low error rates. Yield is dependent upon several factors:
- Read Length: the longer the read, the more data.
- Read Type: paired end reads yield twice the data as single read.
- Optimal Cluster density: it is imperative to accurately quantitate your library to ensure high data yield. We do our best to QC libraries before sequencing, but we cannot pool samples for you.
- High Quality Library: libraries that contain a high level of adapter dimers will yield significantly less data, as the fragments will hybridize to the flowcell. Similarly, over amplified libraries can negatively impact yield.
- Uniform Base composition: libraries that have uneven base composition tend to pose problems with the HiSeq analysis software. These issues can be mitigated using several strategies, but the net effect will be lower data yield per lane than a balanced library.
Please see Illumina’s HiSeq 2500 Specification page for current data yields. While we regularly achieve greater than ‘spec’ yields from the HiSeq2500, typicially matching yields for High Output mode, it is best to be conservative when planning your experiments.
We will also be happy to run your samples in High Output mode, but you will need to submit a full flowcell (ie. 8 lanes/libraries) at a time.
Per lane sequencing: Data will be returned in Sanger FASTQ format via our high speed aspera server
End to end services: alignment files, variant calls, and any intermediate files you wish via aspera server
Whole Genome: pipeline data provided through Illumina Genome Network, returned via hard drive
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