ONT Lite Whole Plasmid Sequencing

• Library preparation (amplification-free, including minimal fragmentation); using the rapid barcoding v14 library prep chemistry from Oxford Nanopore Technologies
• Sequencing using Oxford Nanopore GridION (flow cell R10.4.1) 
• Bioinformatics included: 
  • Comprehensive data analysis report with plasmid map, read histogram, coverage plot, assembly statistics and QC (.HTML)
  • Consensus plasmid sequence (.FASTA) and incl. quality scores (.FASTQ)
  • Annotated plasmid sequence of all contigs in GENBANK format (.GBK) (import directly into your favourite software like Sanger's Artemis, SnapGene, Geneious Prime, Benchling, SeqBuilder Pro etc.)
  • Plasmid map (.PNG, .HTML)
  • Per-base clonal purity data with confidence score (.CSV)
  • Raw plasmid sequence reads that aligned to the assembled plasmid (.FASTQ)
  • Annotated feature table, including the nucleotide sequence of each feature (.CSV)
  • Interested in Pod5: please use our premium service (see below).  Your data will be securely delivered through your Ecom account 

Use the following concentration and volume for your samples:

Please note: Samples that do not meet our concentration requirement might fail in sequencing or lead to inaccurate assemblies (link to FAQ). No repetition will be performed on failed samples due to wrong concentrations adjustment.

*TAT = Turnaround time

Sample Requirements

• Sample Type: Clonal, circular, double-stranded plasmid DNA at the minimum volume and at concentration specified.
• Do not use NanoDrop for DNA quantification. Spectrophotometric methods are unreliable—please use a fluorometric method such as Qubit for accurate concentration measurement.
• Resuspension buffer: Nuclease-free water or elution buffer (10 mM Tris, pH 8.5).
• Purity (OD260/280): 1.8–2.0
• Purity may be analyzed with Nanodrop or other spectrophotometric methods. Avoid ethanol contaminations – if using Qiagen miniprep kit, we recommend performing the optional PB buffer wash step to increase purity.

Format:

• Barcode labelled 1.5 ml Eppendorf Safe-Lock tubes
• Avoid sealing tubes with tape or parafilm
• Label tubes with our free barcode labels and affix them horizontally for easy scanning

Shipment method:

• Use nearest DropBox location for free sample pickup or select a courier service of your choice
• Ensure secure transport by using our approved sample bags and boxes
• Ship all samples to our Eurofins Genomics sequencing lab in Cologne, Germany

The WPS Premium Service guarantees exclusive use of a fresh flow cell dedicated entirely to your samples. To ensure maximum data integrity, the flow cell is discarded immediately after sequencing.

If your project demands strict data purity and zero risk of cross-contamination with other customer samples, we highly recommend choosing our Premium Service. This ensures complete isolation of your sequencing run on a dedicated single-use flow cell, fully eliminating the risk of index hopping (see FAQ section on “Index hopping”).

How can I order the Premium Service ?

The Premium Service is only available for samples submitted in plates. During order placement, please check the "Premium Service" checkbox on the order page. Please note that this service incurs an additional fee.