Zymo Research는 1994년도에 설립된 미국 회사로 고품질 키트 제조회사 입니다.
각종 샘플(세포, 조직, 환경샘플 등)로부터 고품질
DNA나 RNA를 가장 쉽고 빠르게 뽑을 수 있습니다.
이 외에도, 각종 Epigenetics 관련 제품들
(DNA Methylation kit 등)과 Microbiomics
(샘플 채집부터 분석까지의 전 단계의 제품)
연관 제품들이 준비되어 있습니다.
Highlights
Co-Detection of Genetic and Epigenetic Information: Seamlessly analyze both genomic data and DNA methylation in a single experiment, with the validated open-source bioinformatics tools and comprehensive step-by-step guides.
True-End Fragment Analysis: Designed for optimal performance with small or degraded DNA fragments, including cell-free DNA (cfDNA) and FFPE-derived DNA, to capture the fragment’s true end.
Accurate Methylation Detection: Achieve precise methylation calling with a direct ligation-based protocol that preserves native termini, ensuring accuracy in each DNA fragment analysis.
서한형 대리
Zymo Research 제품 담당자
경신과학(주)
영업부
H.P) 010-8832-6303
HanHyung Seo
Zymo Research Brand Manager
Kyongshin scientific Co., Ltd.
Sales Department
H.P) 82)10-8832-6303
제품소개
Highlights
Co-Detection of Genetic and Epigenetic Information: Seamlessly analyze both genomic data and DNA methylation in a single experiment, with the validated open-source bioinformatics tools and comprehensive step-by-step guides.
True-End Fragment Analysis: Designed for optimal performance with small or degraded DNA fragments, including cell-free DNA (cfDNA) and FFPE-derived DNA, to capture the fragment’s true end.
Accurate Methylation Detection: Achieve precise methylation calling with a direct ligation-based protocol that preserves native termini, ensuring accuracy in each DNA fragment analysis.
Product Description
The Zymo-Seq Trio WGBS Library Kit offers an optimized and reliable workflow for whole genome bisulfite sequencing (WGBS) library preparation from cell-free DNA (cfDNA), FFPE DNA, and gDNA. This all-inclusive kit features a straightforward procedure capable of preparing high-quality methyl-seq libraries from as little as 5 ng of cfDNA or fragmented DNA. The process is completed in three basic steps: bisulfite conversion, direct adapter ligation, and index PCR amplification.
The initial bisulfite treatment is gentle on fragmented DNA, such as cfDNA, yet effective in converting any unmodified cytosines into uracil. Next, the innovative splinted adapters capture and directly ligate the Illumina-compatible adapters onto any size DNA fragment, allowing for sequencing of nicked, damaged, and short DNA fragments that may have otherwise been discarded with traditional library prep methods. The direct ligation of the adapters also eliminates the need for second-strand synthesis, end repair, and dA tailing steps, thus reducing bias by preserving the integrity of any native methylation present on the fragment termini. Finally, validated open-source bioinformatic tools and a step-by-step guide are available for customers to co-detect genetic information, enhancing downstream analysis power.
Technical Specifications
Barcode Sequences
Please refer to the Documents Section.
Bisulfite Conversion
>99.5% of non-methylated cytosine residues are converted to uracil; >99.5% protection of methylated cytosines.
Equipment Required
Thermal cycler(s) with temperature adjustable lids, microcentrifuge, magnetic stand.
Hands-On Time
~2 hours
Input Quality
For optimal results, use at least minimum input of purified fragmented DNA with no RNA or gDNA contamination. Fragmented DNA can be concentrated using the DNA Clean & Concentrator (D4013) prior to processing. Input DNA can be suspended in water, DNA Elution Buffer, or TE buffer.
Library Storage
Libraries eluted in DNA Elution Buffer (provided) may be stored at ≤ 4°C overnight or ≤ -20°C for long-term storage.
Maximum Input
10 ng
Minimum Input
5 ng
Sample Input Material
Purified cell-free DNA (cfDNA), Sonicated FFPE DNA (average size of 300-600 bp), Sheared genomic DNA (average size of 300-600 bp)
Sequencing Platform Compatibility
Libraries are compatible with all Illumina sequencing platforms. Recommended: NextSeq, NovaSeq.
It is possible to generate libraries with inputs lower than 5 ng; however, the quality of such libraries is not guaranteed. Typically, at lower inputs the percentage of adapter dimers increases in the final library. For best results, we recommend concentrating the sample input as much as possible to meet the 5 ng minimum input prior to using the kit. Samples can also be concentrated using a DNA Clean & Concentrator Kit and/or during the bisulfite conversion step by reusing the same column for clean-up.
It is possible to generate libraries with inputs higher than 10 ng; however, the quality of such libraries is not guaranteed. For best results, we recommend diluting your sample down to meet the 10 ng maximum input.
We recommend up to 4 freeze-thaw cycles for the Adapter Ligation Buffer 2 (orange cap), Adapter Ligation Buffer 3 (yellow cap), and the Adapter Ligation Master Mix (green cap). After 4 freeze-thaw cycles, the quality of the libraries may vary. We recommend making additional aliquots of these buffers upon first use as necessary to minimize the potential freeze-thaw cycles the reagents undergo. The Adapter Ligation Buffer 1 (red cap) is not as sensitive to freeze-thaw cycles and therefore is not affected by being thawed more than 4 times.
Each tube/well contains a pre-mixed forward and reverse primer set that contain a unique i5 and i7 index, respectively. The concentration of each UDI primer pre-mix is 5 µM total (2.5 µM each primer).
Libraries are suitable for any cycling number, but increased cycling numbers will require greater amounts of adapter trimming for the shorter library fragments. For most applications, 100 base paired-end (PE) reads are enough to generate substantial amounts of high-quality data for genome-wide coverage. The sequencing depth will be dependent on the genome size, genome coverage, and site coverage required. Generally, aiming for 10X coverage per CpG site is recommended.
Using 100 bp PE sequencing, we recommend at least 500 million reads for human WGBS at 10X CpG coverage, and at least 400 million reads for mouse WGBS at 10X CpG coverage.
