White Paper : NGS（Next Generation Sequencer）Application of nanoimprint
lithography to chip mass production
1. Executive Summary

Nanoimprint technology was originally developed about 25 years ago as a fine patternformation technology for semiconductors. In recent years, optical sensors for mobile devices,automatic driving, robotics, security, etc., and light guide plates for augmented reality (AR)glasses have become major applications, and the technology has been reconsidered forstructure formation applications such as DOE and MLA at a level of few hundred nanometers tofew micrometers. The application of NGS discussed here is not for optical components, but fornanoimprint technology as the one for mass-producing substrates with a fine hole structure in
the reading area of DNA testing chips.

2. Introduction

Next-generation sequencers (NGS), which were introduced in the United States in the mid2000s, are devices that can read gene sequences at high speed. The number of DNA
fragments that can be read simultaneously in NGS is greater than in conventional DNA
sequencers, making it possible to analyze genomes (genetic information) at an overwhelmingly
low cost and in a short time. The NGS equipment market is expected to quadruple over thenext 10 years, starting in 2016 (according to SEED Planning data). The current NGS market
has been led in development by Illumina, the largest DNA sequencer company in the US.
The development of sequencers for the "new next generation" is also gaining momentum,especially in the US and China. In contrast to next-generation sequencers, which generallyamplify DNA and read out the base sequence by optically detecting fluorescent labels, this newmethod does not amplify DNA but electrically reads out the base sequence one molecule at atime. In principle, it can read out nucleotide sequences faster and at a lower cost than current
next-generation sequencers.
What can be done with NGS?
First, it can be used to determine nucleotide sequences. For example, genome sequencing
and comprehensive sequencing of transcripts (mRNA). The high performance and low cost of
next generation sequencers has brought them to a level that most people can afford.
Another thing that can be done is to know the quantity. Next-generation sequencers are ableto determine the nucleotide sequence of a variety of DNA in a mixed state. This is because themachine is able to separate the diverse DNA into individual molecules. Therefore, the DNA thatwas more abundant in the DNA used for input will result in more output, and the DNA that wasless abundant will result in less output. As a result, it is also possible to know the frequency ofnucleotide sequences. This feature has led to the development of a variety of applications. For
example, the gene expression levels can be measured.
Metagenomics, which is the sequencing of the genomes of organisms in soil and water, canalso be used to determine what kinds of organisms were present in the soil and to whatextent.
In addition, by examining where transcription factors, such as the Yamanaka 4 factor(famous for iPS cells), bind to genomic DNA, we can determine what genes are involved in theregulation of gene expression. Other methods have also been developed to predict theevolution of organisms over time based on the diversity and frequency of SNPs in the genome.
NGS is having a particularly large impact in the field of cancer medicine, where nextgeneration sequencers are being used to comprehensively analyze cancer gene information,
leading to the development of drugs that target key genetic abnormalities (molecular targeteddrugs). There is a growing movement to introduce this into daily practice rather than research.

SCIVAX | NANOBIC 7-7 Shin-Kawasaki, Saiwai-ku, Kawasaki City, Kanagawa Prefecture 212-0032 | +81-44-599-5051

3. Thesis Questions

In the NGS industry, sequencing became possible at a cost of about US$1,000 a few years
ago now, but there is a need for further cost reduction, faster processing, and accuracy. In thiscontext, the cost of sequencing equipment as well as the cost of testing per specimen, i.e., thecost of chips, can be reduced, thereby expanding the range of applications of NGS, including
clinical applications, and the market is expected to grow.

4. Solutions

NGS devices and their dedicated chips, which are still in the development stage, have theirown unique well structure. The hole structure has usually a diameter of 0.5 to 1um, and theinside of the hole is hydrophilic, while the rest of the hole is hydrophobic, allowing DNA orpolymerase to be added to the hydrophilic part, and DNA input solution to selectively adhere to
the hole part. Usually, it is possible to create similar results using an i-line exposure systemother than nanoimprint for a hole of this size, but nanoimprint is superior in two major ways.
First, when a hole diameter smaller than 0.5um is required, where there are complexities tomold using an i-line lithography system, in the case of nanoimprint, it can be done as long as
the mold can be created using an excimer laser source used for semiconductor lithography.
Second, the process is simple, making it easy to achieve mass production effect while reducing
its cost.

5. Products Info

As a mass production process for NGS chips, SCIVAX has developed a material and process
to create a hydrophilic APTMS hole region surrounded by a hydrophobic HMDS region on aglass wafer and has launched a mass production line. (The process on the left in the figure
below) In addition, for more advanced NGS customers, we are developing a process to formhydrophobic well 3D hole structures on glass wafers using a resin that has no effect on
autofluorescence.

Forming the Hydrophilic hole region
in the Hydrophobic surface withoutleaving well structure.

Forming the Hydrophobic wellstructure on Hydrophilic substratesurface.

6. Conclusion

As described above, nanoimprint technology has been recognized as an essential mass
production technology for the technical and business development of NGS, and SCIVAX has
launched a foundry service for mass production of NGS chips using the most suitableequipment, materials and processes developed in-house.

SCIVAX | NANOBIC 7-7 Shin-Kawasaki, Saiwai-ku, Kawasaki City, Kanagawa Prefecture 212-0032 | +81-44-599-5051