ONT Lite Clonal Amplicons

ONT Lite Clonal Amplicon Sequencing

Clonal / linear amplicon sequencing using Oxford Nanopore Technology.

Long-read clonal / linear amplicon sequencing is a uniquely designed service for scientists looking to perform full-length sequencing and annotation of clonal, linear, double-stranded DNA between 500 bp - 25 kb in length using Oxford Nanopore Technologies. A clonal amplicon is a PCR product derived from a single DNA or cDNA template molecule or a group of genetically identical molecules. Ideally, the PCR amplicon should be specific, resulting in a single distinct band on an agarose gel or a similar quality control outcome. If you have non-clonal amplicons, such as those from non-specific PCR reactions or mixtures, please consider using our Custom Long-Read Amplicon Sequencing service >>.

Take advantage of super low prices of 15€ per sample and exceptionally fast turn-around time of 1-3 days.

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Applications & services

Clonal full-length amplicon sequencing
Amplicon assembly, amplicon map & clonal purity check
Sequencing of GC rich and hairpin regions

Specifications

From one sample
Dropbox shipment possible
Super fast turnaround times
Prepaid option available
Library preparation (amplification-free, including minimal fragmentation); using v14 library prep chemistry
Sequencing using Oxford Nanopore GridION (flow cell R10.4.1)
Raw reads to generate a high-accuracy linear consensus sequence
Bioinformatics analysis always included
Not suited for ssDNA

>> FAQ's clonal amplicons

>> Trouble- shooting

>> how to redeem prepaid coupons

>> Sequencing Accessories

This product is for clonal amplicons only!

A clonal amplicon is a PCR product that originates from a single DNA or cDNA template molecule or a genetically identical group of molecules. The PCR amplicon should be a specific amplicon product, i.e. a single band on an agarose gel or similar QC result. For non-clonal amplicons (e.g. unspecific PCR reactions or mixtures) please use our Custom Long-Read Amplicon Service >>.

Starting material, deliverables, & sample submission

Deliverables

Data analysis report in HTML format
Amplicon assembly (FASTQ and FASTA format) & amplicon feature map
Amplicon Clonal Purity Check; per base Quality with confidence score
Annotated genome sequence of all contigs in GENBANK format (import directly into your favourite software like Sanger's Artemis, SnapGene, Geneious Prime, Benchling, SeqBuilder Pro etc.)
Annoated feature table, including the nucleotide sequence of each feature
Sequencing Reads QC

Your data will be securely delivered through your Ecom account.

No delivery of Pod5 or FASTQ files, that do not align.

If you are interested in Pod5 files and un-aligned FASTQ files please contact us, regarding our premium service.

Starting material

Amplicon size: 500 bp - 25 kb
Sample type: purified PCR products
Purity (OD260/280): 1.8-2.0
Volume: 20µL
Concentration: 30 ng/µl
Minimum Amount: 600 ng
Resuspension Buffer: Water, EB, or low TE (<0.1 mM EDTA)
Format: barcode labelled 1.5 ml safe-lock tubes
Shipment Method: Room temperature

Sample preparation & submission

Sample Preparation & Submission

Use the following concentration and volume for your samples:

Sample type	Length	Concentration	Min. Volume
Linear / clonal amplicons	0.5 kb - 25 kb	30 ng/µl	20 µl

Please note: Samples that do not meet our concentration requirement might fail in sequencing. No repetition will be performed on failed samples due to wrong conc adjustment.

Please send your sample tubes according to the requirements below:

Use 1.5 ml safe-lock tubes for your samples
Please provide your samples either in nuclease-free water or elution buffer (10 mM Tris, pH 8.5)
Please do not use Nanodrop for your concentration measurement. Spectrophotometric techniques are unreliable for the quantification measurement. Please use a fluormetric method, like Qubit.
Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection
Label your sample tubes with our free barcode labels. Affix the label horizontally

How and where to ship your samples:

Use either one of your nearest DropBox for free sample pick up or a courier of your choice
You can convinently use one of our sample shipment options to send your samples
To send your samples to us safely, make use of our sample bags and boxes
Send your samples to our Eurofins Genomics sequencing lab in Cologne, Germany

Please ensure that your sample is free of any contaminants (e.g., ethanol) that could negatively affect your results. A complete list of contaminants can be found here: https://community.nanoporetech.com/contaminants

Literature

Clonal Amplicon Sequencing Demo Report