Review




Structured Review

Oxford Nanopore long-read cdna sequencing
Full-length <t> cDNA sequencing </t> (ONT).
Long Read Cdna Sequencing, supplied by Oxford Nanopore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cdna+sequence/cdna+sequencing/pmc07565673-8-5-2
Average 90 stars, based on 1 article reviews
long-read cdna sequencing - by Bioz Stars, 2026-10
90/100 stars

Images

1) Product Images from "Deletion in the Bardet–Biedl Syndrome Gene TTC8 Results in a Syndromic Retinal Degeneration in Dogs"

Article Title: Deletion in the Bardet–Biedl Syndrome Gene TTC8 Results in a Syndromic Retinal Degeneration in Dogs

Journal: Genes

doi: 10.3390/genes11091090

Full-length  cDNA sequencing  (ONT).
Figure Legend Snippet: Full-length cDNA sequencing (ONT).

Techniques Used: Sequencing, Produced

Expression of retinal marker genes. ( a ) The average expression of marker genes for each retinal cell type in the unaffected BE02 ( a ) and GS01 ( b ) dogs as well as the affected GR02 ( c ), based on cDNA sequencing. ( d ) Relative mRNA expression levels by quantitative RT-qPCR in two different regions (exons 7–8 and exons 13–14) of the TTC8 gene, as well as the retinal marker genes GFAP (macroglial cells), OPN1LW and OPN1SW (cone photoreceptors), RHO (rod photoreceptors), RPE65 (RPE cells), and POU4F1 (retinal ganglion cells) expression in three unaffected dogs (BE02, LR02, GS01) and the affected GR02, normalized to GAPDH and ACTB gene expression. Rod: rod photoreceptors, cone: cone photoreceptors, RGC: retinal ganglion cells, AC: amacrine cells, HC: horizontal cells, BC: bipolar cells, MG: macroglial cells, RPE: retinal pigment epithelium, BE02: unaffected beagle, GS01: unaffected German shepherd, GR02: affected golden retriever, LR02: unaffected Labrador retriever.
Figure Legend Snippet: Expression of retinal marker genes. ( a ) The average expression of marker genes for each retinal cell type in the unaffected BE02 ( a ) and GS01 ( b ) dogs as well as the affected GR02 ( c ), based on cDNA sequencing. ( d ) Relative mRNA expression levels by quantitative RT-qPCR in two different regions (exons 7–8 and exons 13–14) of the TTC8 gene, as well as the retinal marker genes GFAP (macroglial cells), OPN1LW and OPN1SW (cone photoreceptors), RHO (rod photoreceptors), RPE65 (RPE cells), and POU4F1 (retinal ganglion cells) expression in three unaffected dogs (BE02, LR02, GS01) and the affected GR02, normalized to GAPDH and ACTB gene expression. Rod: rod photoreceptors, cone: cone photoreceptors, RGC: retinal ganglion cells, AC: amacrine cells, HC: horizontal cells, BC: bipolar cells, MG: macroglial cells, RPE: retinal pigment epithelium, BE02: unaffected beagle, GS01: unaffected German shepherd, GR02: affected golden retriever, LR02: unaffected Labrador retriever.

Techniques Used: Expressing, Marker, Sequencing, Quantitative RT-PCR, Gene Expression

Related Articles

Sequencing:

Article Title: RNA Sequencing in Disease Diagnosis
Article Snippet: .. Third-generation platforms from companies such as PacBio and Oxford Nanopore now also provide the opportunity to significantly extend sequencing of cDNA molecules with average read lengths ranging from 700 to 5,000 bp and beyond ( , , ). ..

Article Title: An endogenous retroviral element co-opts an upstream regulatory sequence to achieve somatic expression and mobility
Article Snippet: 2X100 bp paired-end sequencing was performed on Novaseq (Illumina). .. For long-read Oxford Nanopore (ONT) cDNA sequencing, we first prepared mRNA with Dynabeads mRNA Purification Kit (Invitrogen #61006) starting from 100 μg of DN-ase digested total RNA, according to the manufacturer’s protocol. .. Supplementary Table 1 provides basic sequencing statistics for all samples. mRNA concentration was quantified with the Qubit RNA Broad Range Assay Kit (Thermo Fisher Scientific) and sample quality was checked on Bioanalyzer (Agilent).

Article Title: Mapping the temporal transcriptomic signature of a viral pathogen through CAGE and nanopore sequencing.
Article Snippet: .. We employed CAGE sequencing on the Illumina platform to determine transcript start sites, alongside long-read direct cDNA sequencing on Oxford Nanopore Technology platform to detect full-length viral transcripts. ..

Article Title: Mapping the temporal transcriptomic signature of a viral pathogen through CAGE and nanopore sequencing.
Article Snippet: .. We employed CAGE sequencing on the Illumina platform to determine transcript start sites, alongside long-read direct cDNA sequencing on Oxford Nanopore Technology plat- form to detect full-length viral transcripts. ..

Article Title: Neuronal cell adhesion molecule (NRCAM) variant defined by microexon skipping is an essential, antigenically distinct, and targetable surface proteoform in high-grade glioma
Article Snippet: .. To determine whether skipping of NRCAM microexons 5 and 19 occurs concurrently and in the context of functional, cap-to-poly(A) transcripts, we performed direct cDNA long-read sequencing using Oxford Nanopore platform Promethion P2 for two DMG PDX samples, 7316-1763 and 7316-1769. ..

other:

Article Title: Defining the Temporal Transcriptomic Landscape of a Viral Pathogen through Nanopore and CAGE sequencing
Article Snippet: We employed cap analysis gene expression sequencing (CAGE-Seq) on Illumina platform to determine the transcript start sites alongside longread direct cDNA sequencing (dcDNA-Seq) on Oxford Nanopore Technology platform to detect full-length viral transcripts.

Article Title: Defining the Temporal Transcriptomic Landscape of a Viral Pathogen through Nanopore and CAGE sequencing
Article Snippet: We employed cap analysis gene expression sequencing (CAGE-Seq) on Illumina platform to determine the transcript start sites alongside long-read direct cDNA sequencing (dcDNA-Seq) on Oxford Nanopore Technology platform to detect full-length viral transcripts.

Article Title: The enhancer RNA, AANCR, regulates APOE expression in astrocytes and microglia
Article Snippet: Traditionally, these experiments have been referred to as RNA-sequencing, but since the RNA is reverse transcribed to cDNA which is then sequenced, we called them cDNA-sequencing (to contrast them to actual RNA-sequencing as in Oxford Nanopore direct RNA sequencing experiments).

Purification:

Article Title: An endogenous retroviral element co-opts an upstream regulatory sequence to achieve somatic expression and mobility
Article Snippet: 2X100 bp paired-end sequencing was performed on Novaseq (Illumina). .. For long-read Oxford Nanopore (ONT) cDNA sequencing, we first prepared mRNA with Dynabeads mRNA Purification Kit (Invitrogen #61006) starting from 100 μg of DN-ase digested total RNA, according to the manufacturer’s protocol. .. Supplementary Table 1 provides basic sequencing statistics for all samples. mRNA concentration was quantified with the Qubit RNA Broad Range Assay Kit (Thermo Fisher Scientific) and sample quality was checked on Bioanalyzer (Agilent).

Functional Assay:

Article Title: Neuronal cell adhesion molecule (NRCAM) variant defined by microexon skipping is an essential, antigenically distinct, and targetable surface proteoform in high-grade glioma
Article Snippet: .. To determine whether skipping of NRCAM microexons 5 and 19 occurs concurrently and in the context of functional, cap-to-poly(A) transcripts, we performed direct cDNA long-read sequencing using Oxford Nanopore platform Promethion P2 for two DMG PDX samples, 7316-1763 and 7316-1769. ..



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Image Search Results


Kaplan-Meier survival plots for A53T α-synuclein transgenic mice, A53T/Kcnn1-3copy mice, and A53T/Kcnn1-6copy mice. Median survival time was extended by 47% for the A53T/ Kcnn1-3copy mice and >100% for the A53T/Kcnn1-6copy mice. The two Kcnn1 transgenes are independent. See text for the different motor phenotypes of the A53T and A53T/Kcnn1-6copy mice.

Journal: bioRxiv

Article Title: Neuronal overexpression of mouse potassium channel subunit Kcnn1 in A53T α-synuclein mice more than doubles median survival time, associated with suppression of phospho-S129 α-synuclein formation

doi: 10.64898/2026.03.09.709927

Figure Lengend Snippet: Kaplan-Meier survival plots for A53T α-synuclein transgenic mice, A53T/Kcnn1-3copy mice, and A53T/Kcnn1-6copy mice. Median survival time was extended by 47% for the A53T/ Kcnn1-3copy mice and >100% for the A53T/Kcnn1-6copy mice. The two Kcnn1 transgenes are independent. See text for the different motor phenotypes of the A53T and A53T/Kcnn1-6copy mice.

Article Snippet: The mouse Kcnn1 cDNA sequence (NM_032397), was PCR-amplified from OriGene Technologies plasmid MR208601, omitting the 3’-terminal Myc-DDK tag sequence.

Techniques: Transgenic Assay

Absence of serine129 phospho-α-synuclein (Psynuclein) immunostaining in 12 mo A53T/Kcnn1-6copy mouse as compared with A53T endstage (9 mo) mouse. A. Location of superior colliculus (purple). Sagittal and coronal cross-sections of brain are shown, to illustrate midbrain dorsal position in the sagittal section and paired left and right structures in the coronal views. B. Sagittal views of superior colliculus, from 20 µm sagittal sections ∼1.2 mm from midline, showing presence of Kcnn1 overexpression in A53T/Kcnn1-6copy 12 month old mouse is associated with absence of α-synuclein disease-associated Psynuclein. Upper panels, A53T endstage 9 month old mouse sagitally sectioned and immunostained to reveal copious Psynuclein (left) and endogenous levels of Kcnn1 (right). Lower panels, absence of Psynuclein associated with overexpression of Kcnn1. Red arrow likely denotes intermediate gray layer. Scale bar, 200 μm.

Journal: bioRxiv

Article Title: Neuronal overexpression of mouse potassium channel subunit Kcnn1 in A53T α-synuclein mice more than doubles median survival time, associated with suppression of phospho-S129 α-synuclein formation

doi: 10.64898/2026.03.09.709927

Figure Lengend Snippet: Absence of serine129 phospho-α-synuclein (Psynuclein) immunostaining in 12 mo A53T/Kcnn1-6copy mouse as compared with A53T endstage (9 mo) mouse. A. Location of superior colliculus (purple). Sagittal and coronal cross-sections of brain are shown, to illustrate midbrain dorsal position in the sagittal section and paired left and right structures in the coronal views. B. Sagittal views of superior colliculus, from 20 µm sagittal sections ∼1.2 mm from midline, showing presence of Kcnn1 overexpression in A53T/Kcnn1-6copy 12 month old mouse is associated with absence of α-synuclein disease-associated Psynuclein. Upper panels, A53T endstage 9 month old mouse sagitally sectioned and immunostained to reveal copious Psynuclein (left) and endogenous levels of Kcnn1 (right). Lower panels, absence of Psynuclein associated with overexpression of Kcnn1. Red arrow likely denotes intermediate gray layer. Scale bar, 200 μm.

Article Snippet: The mouse Kcnn1 cDNA sequence (NM_032397), was PCR-amplified from OriGene Technologies plasmid MR208601, omitting the 3’-terminal Myc-DDK tag sequence.

Techniques: Immunostaining, Over Expression

Stereotactic injection of AAV9-CMV-Kcnn1 virus into the right superior colliculus of an asymptomatic 6 month old A53T mouse leads to overexpression of Kcnn1 in right superior colliculus (upper panel, outlined in light blue), associated with absence of Psynuclein (lower panel outlined in blue) observed after 54 days. Shown in upper and lower panels are separately immunostained neighboring 20 µm cryosections of flash-frozen brain. Upper panel, coronal section of brain showing right superior colliculus with clusters of fluorescent anti-Kcnn1-positive cells in what appear to be the bands of superficial, intermediate, and deep gray layers. Fluorescent cells are also observed beyond the boundaries of the superior colliculus, including crossing the midline and ventrally (see text). Lower panel, almost complete absence of Psynuclein staining in injected right superior colliculus region vs non-injected left (outlined and labeled in blue). (For detail on Psynuclein accumulation outside superior colliculus, see text). Scale bar 250 μm.

Journal: bioRxiv

Article Title: Neuronal overexpression of mouse potassium channel subunit Kcnn1 in A53T α-synuclein mice more than doubles median survival time, associated with suppression of phospho-S129 α-synuclein formation

doi: 10.64898/2026.03.09.709927

Figure Lengend Snippet: Stereotactic injection of AAV9-CMV-Kcnn1 virus into the right superior colliculus of an asymptomatic 6 month old A53T mouse leads to overexpression of Kcnn1 in right superior colliculus (upper panel, outlined in light blue), associated with absence of Psynuclein (lower panel outlined in blue) observed after 54 days. Shown in upper and lower panels are separately immunostained neighboring 20 µm cryosections of flash-frozen brain. Upper panel, coronal section of brain showing right superior colliculus with clusters of fluorescent anti-Kcnn1-positive cells in what appear to be the bands of superficial, intermediate, and deep gray layers. Fluorescent cells are also observed beyond the boundaries of the superior colliculus, including crossing the midline and ventrally (see text). Lower panel, almost complete absence of Psynuclein staining in injected right superior colliculus region vs non-injected left (outlined and labeled in blue). (For detail on Psynuclein accumulation outside superior colliculus, see text). Scale bar 250 μm.

Article Snippet: The mouse Kcnn1 cDNA sequence (NM_032397), was PCR-amplified from OriGene Technologies plasmid MR208601, omitting the 3’-terminal Myc-DDK tag sequence.

Techniques: Injection, Virus, Over Expression, Staining, Labeling

Absence of Psynuclein from right superior colliculus along ∼1 mm of rostro-caudal length of the AAV9-CMV-Kcnn1-injected mouse shown in (Mouse 1). The right and left superior colliculus are outlined in blue in all sections. Note that the shape of the outline is slightly altered from frame to frame to account for its position relative to surrounding landmarks (not shown) and compared with The Mouse Brain in Stereotaxic Coordinates, Franklin and Paxinos, 2007 . Coronal sections shown are spaced every 200 µm rostrocaudally. Consecutive 20 µm coronal sections of brain were taken, beginning at an arbitrary point several 100 µm rostral to superior colliculus and proceeding into the superior colliculus. Every 10 th section (sxn) was stained for Psynuclein. There is absence of Psynuclein from the injected right superior colliculus along the entire cut region. Section #89 appears to contain an artefact in its left aspect. Scale bar 250 μm. Figure 4B. A53T mouse injected with injection solution lacking virus (Control). Sections were collected and immunostained as in A. Superior colliculi circled in blue. There is symmetric presence of Psynuclein in the superior colliculi and outside of it. Scale bar 250 μm.

Journal: bioRxiv

Article Title: Neuronal overexpression of mouse potassium channel subunit Kcnn1 in A53T α-synuclein mice more than doubles median survival time, associated with suppression of phospho-S129 α-synuclein formation

doi: 10.64898/2026.03.09.709927

Figure Lengend Snippet: Absence of Psynuclein from right superior colliculus along ∼1 mm of rostro-caudal length of the AAV9-CMV-Kcnn1-injected mouse shown in (Mouse 1). The right and left superior colliculus are outlined in blue in all sections. Note that the shape of the outline is slightly altered from frame to frame to account for its position relative to surrounding landmarks (not shown) and compared with The Mouse Brain in Stereotaxic Coordinates, Franklin and Paxinos, 2007 . Coronal sections shown are spaced every 200 µm rostrocaudally. Consecutive 20 µm coronal sections of brain were taken, beginning at an arbitrary point several 100 µm rostral to superior colliculus and proceeding into the superior colliculus. Every 10 th section (sxn) was stained for Psynuclein. There is absence of Psynuclein from the injected right superior colliculus along the entire cut region. Section #89 appears to contain an artefact in its left aspect. Scale bar 250 μm. Figure 4B. A53T mouse injected with injection solution lacking virus (Control). Sections were collected and immunostained as in A. Superior colliculi circled in blue. There is symmetric presence of Psynuclein in the superior colliculi and outside of it. Scale bar 250 μm.

Article Snippet: The mouse Kcnn1 cDNA sequence (NM_032397), was PCR-amplified from OriGene Technologies plasmid MR208601, omitting the 3’-terminal Myc-DDK tag sequence.

Techniques: Injection, Staining, Virus, Control

Magnified views of corresponding regions of left and right superior colliculus from an AAV9 CMV-Kcnn1 virus-injected mouse at level of intermediate gray layers, immunostained with anti-Kcnn1 and anti-P-synuclein, showing near complete absence of Psynuclein in the presence of overexpressed Kcnn1. Displayed are 400 µm X 400 µm squares, taken from coronal section #60 (Kcnn1) and section #59 (Psynuclein) of Mouse 1, with their centers ∼1100 microns from either side of the midline. Anti-Kcnn1 (upper panels) shows brighter Kcnn1-immunoreactive cells on the virus-injected right side vs much less bright cells (endogenously expressing Kcnn1) on the uninjected left side. The number of Kcnn1-immunostained cells was roughly equal on the two sides (data not shown). Anti-Psynuclein (lower panels) shows copious intracellular Psynuclein on the left, in the setting of endogenous Kcnn1, and also that Psynuclein extends into processes and that a portion may be in glia or neuropil. On the right, in the setting of overexpressed Kcnn1, only four or five flecks of Psynuclein are visible, the rightmost of which appears to be intracellular. Scale bar 20 µm.

Journal: bioRxiv

Article Title: Neuronal overexpression of mouse potassium channel subunit Kcnn1 in A53T α-synuclein mice more than doubles median survival time, associated with suppression of phospho-S129 α-synuclein formation

doi: 10.64898/2026.03.09.709927

Figure Lengend Snippet: Magnified views of corresponding regions of left and right superior colliculus from an AAV9 CMV-Kcnn1 virus-injected mouse at level of intermediate gray layers, immunostained with anti-Kcnn1 and anti-P-synuclein, showing near complete absence of Psynuclein in the presence of overexpressed Kcnn1. Displayed are 400 µm X 400 µm squares, taken from coronal section #60 (Kcnn1) and section #59 (Psynuclein) of Mouse 1, with their centers ∼1100 microns from either side of the midline. Anti-Kcnn1 (upper panels) shows brighter Kcnn1-immunoreactive cells on the virus-injected right side vs much less bright cells (endogenously expressing Kcnn1) on the uninjected left side. The number of Kcnn1-immunostained cells was roughly equal on the two sides (data not shown). Anti-Psynuclein (lower panels) shows copious intracellular Psynuclein on the left, in the setting of endogenous Kcnn1, and also that Psynuclein extends into processes and that a portion may be in glia or neuropil. On the right, in the setting of overexpressed Kcnn1, only four or five flecks of Psynuclein are visible, the rightmost of which appears to be intracellular. Scale bar 20 µm.

Article Snippet: The mouse Kcnn1 cDNA sequence (NM_032397), was PCR-amplified from OriGene Technologies plasmid MR208601, omitting the 3’-terminal Myc-DDK tag sequence.

Techniques: Virus, Injection, Expressing