GRCF High Throughput Sequencing Center
Our goal is to provide the Johns Hopkins community access to Next Generation sequencing platforms.
We currently feature MiSeq and NovaSeq6000 instruments. We do our best to sequence samples in a timely manner, but our primary focus is on generating high quality data.
High Throughput Sequencing gives you options…
– We offer free consultation.
– We offer simple, per lane and per flow cell pricing, along with end to end service for whole exome/custom targeted projects, RNASeq, and whole genome.
– For NovaSeq 6000 lane loading, we now offer single lane loading on NovaSeq SP to accommodate users who do not need the throughput of a full flowcell.
NovaSeq Per Lane Order
SP: ~800 million paired reads, ~400 million single reads
NovaSeq Per Flow Cell Order
SP: ~1.6 billion paired reads, S1: ~3.2 billion paired reads, S2: ~8.2 billion paired reads, S4: ~20 billion paired reads
HighSeq 2500
We no longer offer HiSeq2500 runs due to cost. Please consider ordering a single lane on NovaSeq. If you have a legacy project that requires HiSeq, please contact David Mohr
Pricing Tables
NovaSeq runs include additional cycles for dual indexed libraries.
If you need to individually load lanes on the NovaSeq, it adds $150/lane
SP: ~1.6 billion paired reads, ~800 million single reads
S1: ~3.2 billion paired reads, ~1.6 billion single reads
S2: ~8.2 billion paired reads, ~4.1 billion single reads
S4: ~20 billion paired reads, ~10 billion single reads
100 cycle: $3600 100 cycle: $6000 100 cycle: $9300
200 cycle: $4300 200 cycle: $7000 200 cycle: $11100 200 cycle: $16000
300 cycle: $4600 300 cycle: $7500 300 cycle: $11600 300 cycle: $17400
500 cycle: $5800
NovaSeq Per Lane Pricing
SP: ~800 million paired reads, ~400 million single reads
100 cycle: $1950
200 cycle: $2300
300 cycle: $2450
500 cycle: $3050
Sample Requirements
For NovaSeq runs, please submit at least 50ul of your sample at 4nM.
Samples should be pooled
Nanodrop is not reliable. Intercalating dye methods or qPCR are preferred.
MiSeq Runs
Please submit 10μl of your sample at 2nM for MiSeq.
Quality Control
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 prior to submission.
Understanding Data Yield
Our facility features the NovaSeq platform. Yield is dependent upon several factors:
Read Length & Read Type
The longer the read, the more data. Paired end reads yield twice the data as single read.
Uniform Base Composition
Libraries that have uneven base composition tend to pose problems with the analysis software. These issues can be mitigated using several strategies, but the net effect will be lower data yield than a balanced library.
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, particularly on the NovaSeq6000. Similarly, over amplified libraries can negatively impact yield
Please see Illumina’s NovaSeq6000 Specification page for current data yields. While we regularly achieve greater than ‘spec’ yields from the NovaSeq6000, it is best to be conservative when planning your experiments.
