Illumina Sequencing Platform

Illumina Sequencing Platform





Sequencing Principle

Illumina sequencing technology, sequencing by synthesis (SBS), is a widely adopted next-generation sequencing (NGS) technology worldwide, responsible for generating more than 90% of the world’s sequencing data. A fluorescently labeled reversible terminator is imaged as each dNTP is added, and then cleaved to allow incorporation of the next base. Since all 4 reversible terminator-bound dNTPs are present during each sequencing cycle, natural competition minimizes incorporation bias. The method virtually eliminates errors and missed calls associated with strings of repeated nucleotides (homopolymers). Illumina sequencing by synthesis technology supports both single-read and paired-end libraries. SBS technology offers a short-insert paired-end capability for high-resolution genome sequencing, as well as long-insert paired-end reads for efficient sequence assembly, de novo sequencing, and more. The combination of short inserts and longer reads increases the ability to fully characterize any genome.

Application


The massively parallel sequencing technology known as next-generation sequencing (NGS) has revolutionized the biological sciences. With its ultra-high throughput, scalability, and speed, NGS enables researchers to perform a wide variety of applications and study biological systems at a level never before possible. For example, NGS allows researchers to:
  • Rapidly sequence whole genomes
  • Zoom in to deeply sequence target regions
  • Utilize RNA sequencing (RNA-Seq) to discover novel RNA variants and splice sites, or quantify mRNAs for gene expression analysis
  • Analyze epigenetic factors such as genome-wide DNA methylation and DNA-protein interactions
  • Sequence cancer samples to study rare somatic variants, tumor subclones, and more Study microbial diversity in humans or in the environment
Today’s complex genomic research questions demand a depth of information beyond the capacity of traditional DNA sequencing technologies. Next-generation sequencing has filled that gap and become an everyday research tool to address these questions.

Actual Data Cases (DNA)

Sample Total base GC% Q20 Q30
Animal A -1401216128 0.4576 0.9736 0.9317
Animal B -1142161250 0.4924 0.9769 0.9339
Animal C -86535604 0.4389 0.9769 0.9364
Crop A -765480092 0.4448 0.975 0.9312
Crop B -922693062 0.4307 0.9749 0.9312
Crop C -1104129616 0.4184 0.9793 0.9382
Fish A 424014960 0.402 0.9814 0.9443
Fish B -196282342 0.3981 0.9755 0.9362
Fruit A -1810084734 0.3961 0.9749 0.9303
Fruit B -1301140888 0.4167 0.9763 0.9326
Fruit C 1794383632 0.4077 0.976 0.9332
Plant A -1381920086 0.3863 0.9715 0.9223
Plant B 999423024 0.4772 0.9758 0.9342
Plant C -255358524 0.5089 0.9803 0.9427
Fungi -632619160 0.4758 0.9777 0.9372

Actual Data Cases (RNA)

Sample Total base rRNA% GC% Q20 Q30
Animal A 81,849,863,700  0.0022 0.4748 0.9773 0.9372
Animal B 76,301,837,700  0.0303 0.5022 0.9788 0.9445
Animal C 69,114,559,500  0.0052 0.5156 0.9783 0.9439
Crop A 938,162,639,100  0.0478 0.4376 0.9801 0.9445
Crop B 644,816,784,000  0.0748 0.5433 0.9792 0.9433
Crop C 388,181,186,700  0.0165 0.5422 0.9806 0.9469
Plant A 501,748,230,600  0.0216 0.4583 0.9797 0.9424
Plant B 296,039,240,400  0.1727 0.5576 0.9759 0.9371
Plant C 257,524,974,750  0.0069 0.4378 0.9768 0.9346
Fruit A 272,597,781,750  0.0295 0.4769 0.9786 0.9397
Fruit B 235,173,258,000  0.0323 0.45 0.9776 0.9357
Fruit C 165,804,670,350  0.0068 0.471 0.9802 0.9435
Fungi A 172,958,883,900  0.0083 0.5836 0.9778 0.9413
Fungi B 158,732,967,900  0.0279 0.5398 0.9822 0.9507
Fungi C 3,558,873,450  0.0042 0.5795 0.9792 0.9471

Successful cases