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CRISPR Mutation Sequencing

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CRISPR Mutation Sequencing

CD Genomics provides CRISPR-Cas9 knockout validation and potential off-target detection in a high-throughput and cost-effective manner by harnessing advanced next generation sequencing (NGS). Our team members have experience in both genome editing and NGS, enabling extensive support to your research goals.

CRISPR-Cas9, which consists of two components: a ‘guide’ RNA (gRNA) and a non-specific CRISPR-associated endonuclease (Cas9), is a new genome editing tool that enables researchers to edit parts of the genome by removing, adding or altering a section of the DNA sequence in any organism. It is currently the simplest, most versatile, precise and effective method of genetic manipulation that has many potential applications including medicine and crop seed enhancement. CRISRP-Cas9 has also been adapted to enable high-throughput genome editing and has revolutionized the generation of targeted mutations.

NGS is a good option to detect the mutations by conducting a PCR using primers flanking the deleted region if you have a large number of samples. In support of the scientific community’s enthusiastic embracement of the CRISPR/Cas9 genome editing technology, CD Genomics developed a cheap, reliable and high-throughput strategy for screening and validating CRISPR-Cas9 based mutations by harnessing amplicon based next-generation Sequencing, which can give you direct and detailed information about the nature and diversity of the mutations, including confirmation of knockout/in allele, assessment of sgRNAs cutting efficiency, homozygous and heterozygous identification, mutation frequencies calculation et al..

CRISPR Mutation Sequencing

One of the challenges in using CRISPR-Cas9 system is that it cuts at a different gene to the one that was intended to be edited, resulting in ‘off-target’, researchers can’t reliably predict when and how often off-targeting occurs. CD Genomics is able to help you to detect the potential off-target effects via targets enrichment and deep sequencing at fairly inexpensive prices. We can sequence hundreds of samples simultaneously. The indel distribution pattern will be analyzed and mutation can be confirmed with the CRISPResso tool.

Features and Benefits:

    • Extensive multiplexing flexibility and high-throughput, sequence up to 108 samples per run
    • Ultra-deep sequencing of amplicons or captured regions. We can provide the sequencing of any targets up to 1,000 bp with >1000X coverage.
    • Highly sensitive detection levels without bias
    • Without laborious and time-consuming cloning steps
    • Dedicated support from specialized Ph.D.-level scientists
  • Cost-effective per sample
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