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KASP Genotyping

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CD Genomics is an experienced (Kompetitive Allele Specific PCR) KASP genotyping service provider, and takes great pride in helping customer every day to obtain high quality genotyping results that advance their research goals.

KASP Genotyping

KASP genotyping system is a homogeneous, fluorescent, endpoint genotyping technology. KASP genotyping assays are based on competitive allele-specific PCR and enable bi-allelic scoring of single nucleotide polymorphisms (SNPs) and insertions and deletions (InDels) at specific loci. Analysis can be carried out in 96-, 384- and 1536-well plate formats. Using KASP genotyping assay, we are able to provide complete genotyping services to our customers. We have a track record of delivering the highest quality data, delivering break-through cost savings across a huge range of project types, and working with hundreds of different species.

Workflow of KASP

Firstly, target-specific KASP primer sequences are designed for each assay, followed by a standard PCR thermal cycle.

KASP PCR reaction uses three components: 1) DNA with SNPs of interest; 2) two different, allele-specific, competing forward primers with unique tail sequence and one reverse primer; 3) FRET cassette plus Taq polymerase.

In the first round of PCR, one of the allele-specific primers matches the target SNP and, with the common reverse primer, amplifies the target region. In the PCR round 2, complement of allele-specific tail sequence is generated. In further rounds of PCR, levels of allele- specific tail increase. The fluor labelled part of the FRET cassette is complementary to new tail sequences and binds, releasing the fluor from the quencher to generate a fluorescent signal. Following completion of the KASP PCR, reaction plates are read in a FRET-capable plate reader and the data analyzed using any cluster analysis viewing software. Detected signals are plotted as a graph, with samples of the same genotype clustering together. The workflow is demonstrated in Figure 1.

Workflow diagram of KSP genotyping procedure Figure 1. Workflow diagram of KSP genotyping procedure.

Sample Requirement

The quantity of DNA that is required is dependent on the scale of your genotyping project and upon the genome size of your study organism. The minimum mass of DNA that we accept per sample is 2 ng, and the minimum concentration that we accept per sample is 5 ng / μl. DNA samples should be submitted in 96- or 384- well plates.

Key Features and Advantages

  • Extensive experience. Have worked with more than 100 individual genotyping projects on ~10 different species.
  • Supporting life science researchers in diverse application areas. Include crop and livestock development, healthcare and diagnostics and et al.
  • Strict quality control. Include quality control of the initial DNA samples, negative and positive control in the sample processing batch to exclude DNA contamination and ensure a technically correct laboratory process.
  • Cost effective. More affordable than other SNP genotyping approaches when the project contain small number of SNPs and large number of samples.
  • High accuracy. >99.8% based on independent assessment.
  • Tremendous flexibility. Flexible primer design, support low to medium throughput study, compatibility with diverse thermal cyclers and plate readers.

Workflow diagram of KSP genotyping procedure

The KASP is a uniplex SNP genotyping platform, it is suitable to be used when there are many SNPs in a few samples or when there are few SNPs in many samples.

Our KASP technology is a simple, cost-effective and flexible system enabling delivering extremely high levels of assay with robustness and accuracy. Our KASP genotyping system has been used successfully in a wide variety of organisms, achieving well over 90% SNP to assay conversion rate.

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CONTACT CD GENOMICS

45-1 Ramsey Road, Shirley, NY 11967, USA
Tel: 1-631-275-3058
Fax: 1-631-614-7828
Email: info@cd-genomics.com