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MALBAC

技术概述

MALBAC(Multiple Annealing and Looping Based Amplification Cycles)多次退火环状循环扩增技术,利用独特的具有链置换活性的DNA聚合酶进行准线性的全基因组预扩增,再通过PCR技术进行指数式扩增,为下游分析提供充足的实验材料。

原 理

不同于以往的非线性或指数型扩增方法,MALBAC技术利用特殊引物,使得扩增子的结尾互补而成环,从而很大程度上防止了DNA的指数性扩增。

MALBAC

技术优势



扩增成功率:扩增成功率高于96%

均一性好:准确进行拷贝数筛查

覆盖度高:全面检测各种单基因病

脱扣率低:准确检测单基因病致病位点





右图是MALBACTM单细胞扩增剂盒与市面上常见的其他四种单细胞扩增剂盒在等位基因脱口率、扩增均一性以及覆盖度方面的比较。图中S、Q、YiKon、R单细胞扩增试剂盒,其原理分别为DOP-PCR,MDA,MDA,MALBAC,MALBAC-like。 从右图中可见:YiKon的MALBAC单细胞扩增试剂盒的等位基因脱扣率(ADO)明显低于其他四种试剂盒。


图为5种单细胞扩增试剂盒等位基因均一性比较

从上图可见:YiKon的MALBAC单细胞扩增试剂盒产物基因拷贝数变异(CNV)的CV值明显小于其他四种试剂盒,其扩增均一性是最好的






从左图可见:在相同测序深度下,MALBAC的基因组覆盖度相比MDA、WGA4有明显优势


取得成果

在国际顶尖杂志发表文章:Science 、Cell 、PNAS等

Genome-Wide Detection of Single-Nucleotide and Copy-Number Variations of a Single Human Cell. Science, 2012, 338, 1622.

Probing Meiotic Recombination and Aneuploidy of Single Sperm Cells by Whole-Genome Sequencing. Science, 2012, 338(6114, 1627-1630)

首次完成人类单个卵细胞的测序

Genome Analyses of Single Human Oocytes. Cell, 2013, 155: 1492-1506

Reproducible Copy Number Variation Patterns among Single Circulating Tumor Cells of Lung Cancer Patients. PNAS, 2013, 110(52): 21083-21088


完成迄今覆盖度最高的单精子基因图谱

Probing Meiotic Recombination and Aneuploidy of Single Sperm Cells by Whole-Genome Sequencing. Science, 2012, 338(6114, 1627-1630)


MALBAC-NGS用于卵裂期胚胎PGS

Validation of Multiple Annealing and Looping-based Amplification Cycle sequencing for 24-chromosome aneuploidy screening of cleavage-stage embryos. Fertility and Sterility, 2014, 102: 1685-1691


MALBAC-NGS用于囊胚PGS

Validation of a next-generation sequencing-based protocol for 24-chromosome aneuploidy screening of blastocysts. Fertility and Sterility, 2016, in press


Live Births after Simultaneous Avoidance of Monogenic Diseases and Chromosome Abnormality by Next-Generation Sequencing with Linkage Analyses. PNAS, 2015, 112(52):15964-9


MALBAC-NGS用于单基因病PGD

A New Next-generation Sequencing-based Assay for Concurrent Preimplantation Genetic Diagnosis of Charcot-Marie-Tooth Disease Type A1 and Aneuploidy Screening. JGG, 2016, Available online 21 January 2016

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