Research ID
hum0612-v1Release info
Research title
Registered research projects for new genetic test analysis data in the restricted-access database
Research overview
- Aims
- The current genetic testing methods are exon capture sequencing and MLPA and limited to detecting point mutations in exons, as well as copy number of exons. We have developed and published papers on novel testing methods, such as vLAS (very long amplicon sequencing), rLAS (RNA long amplicon sequencing) and TgCap (targeted genome capture sequencing), which can detect structural abnormalities, deep intronic and mosaic variants. This study aims to ensure that personal information is handled appropriately when reporting cases with new findings or the development of new genetic testing methods. To this end, we will register these data with the NBDC Human Database for restricted access, thereby facilitating verification and secondary use by other researchers.
- Methods
- vLAS: We amplified peripheral blood DNA using a long PCR set of approximately 20 kb, which covered the entire genomic region, including the introns of the target gene. The amplicons were then pooled and libraries were prepared using Nextera (Illumina), after which they were sequenced on a MiSeq (Illumina).
rLAS: Peripheral mononuclear cells were isolated from whole blood. Total RNA was extracted using Trizol and single-stranded cDNA was synthesized using oligo-dT primers. The full-length mRNA of the ERCC4 gene was then amplified by Long RT-PCR using this as a template. A library was constructed from the amplification products using Nextera (Illumina) technology, and the library was sequenced on a MiSeq (Illumina).
The variant was identified using vLAS on peripheral blood DNA. The splicing abnormality was quantitatively analyzed using rLAS on total RNA from peripheral mononuclear cells. - Participants/materials
- Xeroderma pigmentosum: 1 case (one vLAS specimen and one rLAS specimen)
A novel splicing variant, NC_000016.10(NM_005236.3):c.207+196T>A, was identified in a deep intron of the ERCC4 gene and the patient was diagnosed with type F.
Datasets
| Cart | Dataset ID | Type of data | Analysis method | Access criteria | Date published |
|---|---|---|---|---|---|
| JGAD001129 | NGS (Targeted DNA-seq, Targeted RNA-seq) |
| Controlled-access (Type I) |
Data provider
- Principal investigator
- Yo Niida
- Affiliation
- Division of Genomic Medicine, Department of Advanced Medicine, Medical Research Institute, Kanazawa Medical University
Research projects
No research projects.
Grants
| Name | Title | Project number |
|---|---|---|
Kanazawa Medical University | NA |
|
Related publications
| Title | DOI | Dataset ID |
|---|---|---|
A case with Xeroderma Pigmentosum Type F manifested a mild phenotype due to a deep intronic variant of the ERCC4 gene |
Controlled access users
No use of the controlled access data has been recorded.