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VariantBaits? 液相捕獲定制化Panel

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平臺簡介

VariantBaits?液相雜交捕獲系統可以對客戶感興趣的基因組目標區域進行富集,并在Illumina、Ion Torrent和BGI等二代測序平臺進行高通量測序。通過LC Sciences自主專利的μParaflo®微流體芯片大規模合成高質量寡核苷酸,從而制備超長RNA探針,對帶有測序接頭的基因組文庫進行液相雜交,與目標區域序列互補配對的探針在雜交時特異性結合目的片段DNA,通過鏈霉親和素磁珠與探針上的生物素標記結合,從而抓取并富集目的片段DNA。可用于從小到大規模的基因捕獲,容忍某些局部模板的變異。

LC Sciences采用光原位合成原理,研發出以硅晶板為基礎的大規模合成芯片平臺μParaflo® 微流體芯片,是全球少數幾家擁有此項技術的公司。通過微流控技術對核酸原料進行控制,經由控制光的封閉與否對合成的DNA反應進行控制,達到合成DNA的目的。相比于其他方式,此種DNA合成方法可以實現更大規模的合成以及更可控的DNA合成質量。在合成DNA時,當長度小于1000bp時,芯片合成成本遠低于其他合成方式。在大規模合成DNA的領域當中,芯片合成占有絕對優勢,也是主流選擇。


技術優勢

1 美國LC Sciences探針合成平臺15年技術沉淀,30余篇論文支持
2 120nt超長RNA捕獲探針覆瓦式覆蓋目標區域,擁有更高的錯配容忍度和捕獲效率
3 獨特的探針設計優化,充分利用探針與捕獲區域的結合特性,減少冗余產生
4 可按客戶需求個性化定制Panel,定制周期30個工作日


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合作成果

1.Gao X, Church, GM, et al. (2004) Accurate multiplex gene synthesis from programmable DNA chips. Nature 432, 1050-1054.

2.Porreca GJ, Zhang K, Li JB, Xie B, Austin D, Vassallo SL, LeProust EM, Peck BJ, Emig CJ, Dahl F, Gao Y, Church GM, Shendure J. (2007) Multiplex amplification of large sets of human exons. Nat Methods 4(11), 931-36.

3.Gnirke A, Melnikov A, Maguire J, et al. (2009) Solution hybrid selection with ultra-long oligonucleotides for massively parallel targeted sequencing. Nat Biotechnol 27, 182–89.

4.Lira Mamanova, Alison J Coffey1, et al. (2010) Target-enrichment strategies for nextgeneration sequencing. Nat Methods 7(2):111-8.

5.Teer JK, Bonnycastle LL, et al. (2010) Systematic comparison of three genomic enrichment methods for massively parallel DNA sequencing. Genome Res 20(10), 1420-31.

6.Matzas M, St?hler PF, et al. (2010) High-fidelity gene synthesis by retrieval of sequence-verified DNA identified using high-throughput pyrosequencing. Nat Biotechnol 28(12), 1291-94.

7.Nautiyal S, Carlton VE, Lu Y, Ireland JS, Flaucher D, Moorhead M, Gray JW, Spellman P, Mindrinos M, Berg P, Faham M. (2010) High-throughput method for analyzing methylation of CpGs in targeted genomic regions. Proc Natl Acad Sci 107(28), 12587-92.

8.Myllykangas S, Buenrostro JD, et al. (2011) Efficient targeted resequencing of human germline and cancer genomes by oligonucleotide-selective sequencing. Nat Biotechnol 29, 1024-27.

9.Diep D, Plongthongkum N, Gore A, Fung H, Shoemaker R, Zhang K. (2012) Library-free methylation sequencing with bisulfite padlock probes. Nature Methods 9(3), 270-2.

10.Labrie V, Buske OJ, Oh E, Jeremian R, Ptak C, Gasiūnas G, Maleckas A, Petereit R, ?virbliene A, Adamonis K et al.(2016) Lactase nonpersistence is directed by DNA-variation-dependent epigenetic aging. Nature Structural &Molecular Biology 23(6):566-73.

11.Zaccai F, C T, Savitzki D, Zivony-Elboum Y, Vilboux T, Fitts EC, Shoval Y, Kalfon L, Samra N, Keren Z. (2017) Phospholipase A2-activating protein is associated with a novel form of leukoencephalopathy. Brain 140(2), 370-386.


 

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