ChIP-Seq Library Prep Kit (illumina and MGI Platforms)
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The ChIP-Seq Library Prep Kit (illumina and MGI Platform) was developed for the construction of high quality ChIP-Seq libraries using 5 ng to 400 ng of ChIP DNA as input. The kit is compatible with ChIP DNA fragments generated from both enzymatic methods and physical methods (sonication, nebulization etc.).
Detail
The ChIP-Seq Library Prep Kit (illumina and MGI Platform) was developed for the construction of high quality ChIP-Seq libraries using 5 ng to 400 ng of ChIP DNA as input. The kit is compatible with ChIP DNA fragments generated from both enzymatic methods and physical methods (sonication, nebulization etc.).
ChIP-Seq Library Prep Kit Workflow
ChIP-Seq is the combination of chromatin immunoprecipitation (ChIP) with next generation sequencing. It is a powerful tool for the analysis of global transcription factors and other proteins in diseases and biological pathways, and characterization of histone modifications in a genome-wide level at single-base resolution. ChIP-Seq delivers whole genome level of functional profiling of global transcription factors, and provides better understanding of epigenetic modifications.
Three index types are available for the ChIP-Seq Library Prep Kit of the illumina platform:
Non-index (Cat.# 30032): Libraries do not have index.
Index (Cat.# 30034): Each index primer contains a unique 6-base index sequence can be used for identification. 48 samples can be pooled together. Index information can be downloaded here.
Unique dual index (Cat.# 30036): The ChIP-Seq library multiplexing for 96 samples is possible. Our unique 4-Base Difference Index System have 8 bases index length and at least 4 bases are different from each other for better library identification. Our unique dual indexing primers remove sequencing errors such as index hopping, index contamination, mis-assignment, and other errors. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34034).
Kit advantages:
Super fast protocol
Library prep can be done in 1.5 hours
The hands-on time is only around 10 minutes
Easy procedure
Ready-to-use master mix simplified the procedure
Less reaction components make it easy to setup reactions
Reduced more than half of the beads cost
Input ChIP DNA: From 5 ng to 400 ng
Comparison of library conversion efficiency under the same condition. Input DNA amounts are 5 ng and 30 ng. BioDynami ChIP-Seq Library Prep Kit (Cat.# 30034) was used.
Comparison of aligned reads, aligned rate and duplication rate. Input DNA amounts are 5 ng and 30 ng. BioDynami ChIP-Seq Library Prep Kit (Cat.# 30034) was used.
Data comparison: Input DNA amounts are 5 ng and 30 ng. BioDynami ChIP-Seq Library Prep Kit (Cat.# 30034) was used. Sequencing peak regions are shown.
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HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).This polymerase is also available as a full-length Taq DNA polymerase with a nuclease domain, featuring 100% compatibility with hydrolysis probes (TaqMan® probes etc.).Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.Several independently conducted studies show that HiDi® DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more) For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
Casestudies: HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)
Example Primer Design
Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.
Example Results – There´s no accounting for taste
Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)
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Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.
PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.
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