HiSeq 2000 / 2500

1. 平台简介      

Hiseq 2000测序系统是Illumina公司于2010年推出的一款测序仪,其测序原理与Genome Analyzer II测序系统相似,仍采用可逆终止法的边合成边测序技术。该技术通过将基因组DNA的随机片断附着到光学透明的表面,这些DNA片断通过延长和桥梁扩增,形成了具有数以亿计cluster的Flowcell,每个cluster具有约1000拷贝的相同DNA模板,然后用4种末端被封闭的不同荧光标记的碱基进行边合成边测序。这种新方法确保了高精确度和真实的一个碱基接一个碱基的测序,排除了序列方面的特殊错误,能够测序同聚物和重复序列。 


2. 应用领域      

Hiseq 2000测序系统目前已广泛应用于生物学研究领域,主要包括全基因组de novo测序重测序RNA测序宏基因组测序外显子组测序ChIP-Seq甲基化测序等。


 3. 技术特点       

通量高,单次运行最高可产生600G的数据通量。         
        序列读长相对454较短,可达到100bp。        
        成本较低,操作简易。
 


4. 数据产出

HiSeq 2000

读长

单流动槽的运行时间

双流动槽的运行时间

双流动槽的产量

1×35 bp

~1.5天

~2天

95-105 Gb

2×50 bp

~4.5天

~5.5天

270-300 Gb

2×100 bp

~8.5天

~11天

540-600 Gb

 

HiSeq 2500

测序类型

读长

数据量(lane)

数据量(run)

双端(PE)

2×100   bp

30 Gb

120 Gb

双端(PE)

2×150   bp

45 Gb

180 Gb


5. 实验流程

6. Macrogen应用此平台的优势          

30台HiSeq 2000;          
        Illumina Genome Network全球三家成员之一,同时受 Illumina Genome Network 和 Illumina CSPro 监管,严格使用原厂试剂,是HiSeq 2000平台全球最高质量的代表;          
        平均大于99%碱基准确度达到Q20,保证大于85%碱基准确度达到Q30,平均clean data占raw data90%以上。 


7. Macrogen参与发表文章

Recombination mapping using Boolean logic and high-density SNP genotyping for exome sequence filteringMolecular    Genetics and Metabolism, 2012. 
Exome sequencing identifies null and null as susceptibility genes for monomelic amyotrophyNeuromuscular    Disorders, 2012.    
A homozygous frameshift mutation of sepiapterin reductase gene causing parkinsonism with onset in childhood    Parkinsonism and Related Disorders, 2012. 
Exome sequencing and subsequent association studies identify five amino acid-altering variants influencing human height.    Hum Genet, 2012. 
A conserved two-component regulatory system, PidS/PidR, globally regulates pigmentation and virulence-related phenotypesnull    of Burkholderia glumaeMolecular Plant Pathology, 2012. 
Stabilization of RNT-1, the RUNX homolog of Caenorhabditis elegans, by oxidative stress through a MAP kinase pathway    J Bio Che, 2012. 
Complete mitochondrial genome of Concholepas concholepas inferred by 454 pyrosequencing and mtDNA expression in two mollusc populationsComparative Biochemistry and Physiology, 2012.

A transforming null and null gene fusion in lung adenocarci noma revealed from whole-genome and transcriptome.    Genome Res, 2011. Genetic diagnosis of Duchenne and Becker muscular dystrophy using next-generation sequencing technology:    comprehensive mutational search in a single platform

J Med Genet, 2011.

A transforming null and null gene fusion in lung adenocarci noma revealed from whole-genome and transcriptome.    Genome Res, 2011.
Extensive genomic and transcriptional diversity identified through massively parallel DNA and RNA sequencing of eighteen .    Nature Genetics , 2011.
Discovery of common Asian copy number variants using integrated high-resolution array CGH and massively parallel DNA sequencingNature Genetics, 2010. 
A highly annotated whole-genome sequence of a Korean individualNature, 2009. 
Deletion Hotspots in null Promoter CpG Island Are null-Regulatory Elements Controlling the Gene Expression in the Gene Expression in the ColonPLoS Genet, 2009.



2017年01月04日

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