The DNA Core Facility operates an Oxford Nanopore Technology (ONT) GridION Mk1 offering the ability to sequence a variety of sample types (i.e., genomic DNA, full-length cDNA, PCR amplicons, etc.). ONT is a third generation technology that provides long reads using a novel detection approach. Unlike short read technology, the maximum length of a read is solely dependent on the prepared DNA fragment. Average read lengths of >15 kb are routine. The GridION is capable of processing up to 5 flow cells independently at one time. Typical data yield from a single flow cell is ~8-10Gb in 48 hours (gDNA sample). Throughput is dependent on quality of the sample and, therefore, can be variable.
The facility offers free consultation for researchers in the grant application process or project initiation phase. Staff can discuss current technology capabilities, prepare cost estimates, and provide letters of support. Additional information on technology and applications can be obtained from the Oxford Nanopore website.
Fees for all Oxfordford Nanopore GridION services are listed on the DNACF Fee List
Contact the DNA Core for additional information on conducting a long read sequencing project with the facility.
Submission Guidelines for Sample Type
- General Comments
- Recommended input DNA quality:
- OD 260:280 >1.8
- OD 260:230 between 2.0-2.2 (lower value suggests contaminants; consider re-purification of the sample)
- The concentration of the input DNA should be based on a Qubit measurement (Nanodrop concentrations are unacceptable).
- An image of 100ng of genomic DNA following agarose gel electrophoresis (0.7%) and ethidium bromide staining will be provided by the researcher to demonstrate gDNA integrity.
- Low binding microcentrifuge tubes (0.5 or 1.5mL) are preferred for storing high quality nucleic acids.
|Library Kit||Input||Minimum Concentration Required||Concentration||Recommended
|Oxford Nanopore DNA Library||gDNA||1000 - 1500 ng||10 - 50 ng/ul||EB buffer; 10mM Tris-Cl (pH 8.5)||8-10 Gb; ~1 million reads|
|Oxford Nanopore DNA Library||PCR Amplicon||200 ng||40- 200 ng/ul||EB buffer; 10mM Tris-Cl (pH 8.5)||5-10 million reads|
|Oxford Nanopore cDNA Library||Full Length cDNA||200 ng||5 - 20 ng/ul||EB buffer; 10mM Tris-Cl (pH 8.5)||5-10 million reads|
|Oxford Nanopore 16S Library||gDNA||10 ng||1 ng/ul||EB buffer; 10mM Tris-Cl (pH 8.5)||Full-length 16S gene (~1.5 kb); 5-10 million reads|
|Oxford Nanopore Direct RNA Library||poly A+ RNA||at least 500 ng||50-100 ng/ul||Nuclease-free water||~1 million full-length reads per flow cell|