HEAT-Seq Choice Designs

Overview

HEAT-Seq Choice enables custom designs of between 140 and 20,000 probes. Probes are selected from our database of empirically tested and balanced probes targeting hundreds of oncogenes and thousands of genes associated with human inherited disease.

Benefits

  • Inherited disease and oncology content: The HEAT-Seq Choice database contains probes targeting both strands of nearly all of the coding regions of thousands of genes, enabling straightforward creation of custom designs based on your genes or disease areas of interest. 
  • Simple and easy custom design creation: Custom designs are made using our online design software, NimbleDesign. Simply input your genes or coordinates and NimbleDesign will guide you in making a panel that best meets your needs. 
  • Small to large designs in one tube: HEAT-Seq Choice allows researchers to build custom designs of between 140 and 20,000 probes.

Why use a database?
The design and refinement of custom amplicon designs can be quite challenging. This is particularly true for the approaches based on Molecular Inversion Probes (MIPs), where the advantages of an easy workflow and high precision can be outweighed by the challenges of designing probes that work reliably and provide uniform performance.

Using our expertise and proprietary technology, we have built databases that make the customization process easier. Drawing probes from our databases enable you to spend more time on your research and less time worrying about poor uniformity, dead or poison probes, or creating the probe pools from many individual oligos.

Custom HEAT-Seq Choice designs are made using NimbleDesign, easing the process of getting a high-performance custom panel online.

The power of empirical probe balancing

The performance of HEAT-Seq designs is dramatically improved thanks to proprietary empirical probe performance information.

HSC Data 07-Jul-2016

Figure 1: Sequencing coverage and uniformity improvement from probe balancing

Sequencing coverage is plotted above for equivalent designs made with probes at equimolar ratios (blue) and using the HEAT-Seq Choice product (green). Designs with probes in equimolar concentration can lead to low uniformity and reduced overall coverage. The proprietary HEAT-Seq Choice probe balancing method helps enable a higher level of uniformity and coverage. In this example, uniformity for the entire design was 78.1% and mean coverage was 31.0 X. The equivalent design made using HEAT-Seq Choice systems, whose probe concentrations were adjusted by the proprietary automated process to account for their properties, provides greatly informed uniformity (88.2%) and mean coverage (66.1x). The specific region shown above is of a portion of the MEF2C gene, associated with developmental disorders. The entire custom HEAT-Seq design was made targeting the coding region of 500 genes associated with human genetic or inherited disease.

HSQ Choice Figure 2

 

Figure 2: Sequencing coverage

With equal amounts of sequencing (5M reads in this experiment), the optimized design provides significantly better coverage depth over targets than the design with the probes in equimolar concentrations. This means sequencing resources are used more efficiently and can generate more variant calls. 

Consistent Uniformity

HEAT-Seq custom designs offer high uniformity even with large designs. 

HSQ Choice Uniformity v2

Figure 3: The uniformity of various designs made using the HEAT-Seq Choice database.

Leveraging the empirical data about the performance of probes enables the rapid configuration of designs with reliable uniformity. In the figure above, the uniformity of designs of a variety of sizes is plotted. High uniformity is achieved regardless of design size, which can be exceedingly difficult for other methods. Uniformity is defined is the percentage of probes giving coverage that is greater than or equal to 20% of the mean. 

HEAT-Seq Choice System Performance

 

Custom Design Performance

HEAT-Seq Choice designs provide consistent high performance in the metrics that define high quality data. Create a custom design using your genes or coordinates of interest and proceed to sequencing with confidence. 

HSQ Choice Figure 4

Figure 4: In a custom design made using HEAT-Seq Choice systems targeting 500 genes, the quality of the sequencing data helps to enable highly accurate and complete detection of variants. High uniformity and on-target rate pair with a high level of probe activity to deliver excellent overall coverage, enabling such complete and accurate variant detection. Sequencing for each replicate was subsampled to 5 million reads. Uniformity is defined is the percentage of probes giving coverage that is greater than or equal to 20% of the mean. 

Product Overview

Product Design Size Sizes Available Catalog Number

HEAT-Seq Choice

140-20,000 probes

96 Rxn 7768648001
384 Rxn 7768656001
1536 Rxn 7768664001

Please Contact Us for ordering and questions

For life science research only. Not for use in diagnostic procedures. Unless otherwise stated in references, all data on file.

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