BisCap® methyl hybridization capture sequencing technology is a targeted capture sequencing technology that combines methylation detection with probe hybridization. It can be used to detect targeted regions of DNA methylation of FFPE and cfDNA samples, and is suitable for research of early tumor screening, tumor prognostic marker detection, methylation marker screening and others.
BisCap® methyl hybridization capture sequencing technology uses the post-bisulfite strategy, that is, WGBS library is firstly constructed, and then the targeted library is obtained through the hybridization capture steps, after which sequencing is proceeded by using NGS platforms such as Illumina and MGI. The post-BS strategy avoids the loss of template diversity and is suitable for samples with low amount or poor integrity such as cfDNA and FFPE samples.
In BisCap® panels, the probe design strategy of dual-strand dual-methylation status in CG sites is used for gene regulatory regions (CGI, 5' UTR, Promoter, Silencer, etc.), which ensures methylation detection rates while reducing the probe numbers and total costs.
BisCap®️ methyl hybridization capture sequencing technology is developed based on the proven and simple TargetSeq®️ technology. The BisCap® Enhancer can effectively improve the capture rate, uniformity, and stability.
There are significant differences in DNA methylation status between normal and tumor cells. Changes of methylation status can be detected in early stage of tumorigenesis, and thus DNA methylation is significantly tissue-specific and can be used for tissue tracing. BisCap® panels can be used to detect DNA methylation level and haplotype information in the target regions under single-base resolution and high depth sequencing, which is cost-effective and suitable for methylation marker screening, tumor early screening and other applications.
iGeneTech Bioscience provides both catalog and customized panels with reagent kits and automation upgrade solutions for the whole BisCap® workflow.
| Library Preparation Kits | Hybridization Capture Kits | Target Probes | Equipments |
Of course. We offer customization and semi-customization options to our customers to build their own BisCap® methylation panels.
No. Although the sequences of the adapters are the same, the cytosines in the adapter used in methylation library construction are methylated. They are different products with different names and catalog numbers. Please use the correct version of adapters in methylation library construction.
We use a dual-strand dual-methylation status probe design strategy to design the BisCap® methylation panels. Dual-strand strategy: the methylation statuses of positive and negative strands are normally different, and the dual-strand strategy ensures the capture of both strands. Dual-methylation status strategy: data has shown that, normally, the methylation status of a DNA sequence with continuous cytosines is full-methylation or non-methylation. Therefore, the probes are designed for both these statuses and it is has been proven that this dual-methylation status design strategy is capable of capture most of the targets while reducing the cost hugely compared to the exhaustive strategy.
There are several reasons that might cause low conversion rate of methylated cytosine: 1) The converted library is contaminated by the unconverted library. 2) The bisulfite conversion kit has a short shelf life and the use of an expired kit will significantly affect the result. 3) The input amount is too high.
Pre-BS and post-BS are strategies of adding adaptors before or after BS conversion. BS conversion causes damage to the DNA sequences and some of the DNA will become ssDNA, which leads to a low utilization rate of raw molecules and a much higher input amount requirement.
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