Microfluidic Whole Genome Amplification Device for Single Cell Sequencing

2014 ◽  
Vol 86 (19) ◽  
pp. 9386-9390 ◽  
Author(s):  
Zhilong Yu ◽  
Sijia Lu ◽  
Yanyi Huang
Keyword(s):  
Single Cell ◽  
Whole Genome Amplification ◽  
Whole Genome ◽  
Genome Amplification ◽  
Single Cell Sequencing

scholarly journals The comparison of the performance of four whole genome amplification kits on ion proton platform in copy number variation detection

2017 ◽  
Vol 37 (4) ◽  
Author(s):  
Xinyi Zhang ◽  
Bo Liang ◽  
Xiaoyan Xu ◽  
Feifei Zhou ◽  
Lingyin Kong ◽  
...  
Keyword(s):  
Copy Number Variation ◽  
Single Cell ◽  
Copy Number ◽  
Whole Genome Amplification ◽  
Whole Genome ◽  
Genome Amplification ◽  
Single Cell Sequencing ◽  
Ion Proton ◽  
Number Variation ◽  
Copy Number Variation Detection

With the development and clinical application of genomics, more and more concern is focused on single-cell sequencing. In the process of single-cell sequencing, whole genome amplification is a key step to enrich sample DNA. Previous studies have compared the performance of different whole genome amplification (WGA) strategies on Illumina sequencing platforms, but there is no related research aimed at Ion Proton platform, which is also a popular next-generation sequencing platform. Here by amplifying cells from six cell lines with different karyotypes, we estimated the data features of four common commercial WGA kits (PicoPLEX WGA Kit, GenomePlex Single Cell Whole Genome Amplification Kit, MALBAC Single Cell Whole Genome Amplification Kit, and REPLI-g Single Cell Kit), including median absolute pairwise difference, uniformity, reproducibility, and fidelity, and examined their performance of copy number variation detection. The results showed that both MALBAC and PicoPLEX could yield high-quality data and had high reproducibility and fidelity; and as for uniformity, PicoPLEX was slightly superior to MALBAC.

scholarly journals Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization

10.3791/56394 ◽  
2018 ◽  
Author(s):  
Shukun Chen ◽  
Amin El-Heliebi ◽  
Julia Schmid ◽  
Karl Kashofer ◽  
Zbigniew T. Czyż ◽  
...  
Keyword(s):  
Single Cell ◽  
Target Cell ◽  
Whole Genome Amplification ◽  
Whole Genome ◽  
Genome Amplification

scholarly journals Combined Molecular Genetic and Cytogenetic Analysis from Single Cells after Isothermal Whole-Genome Amplification

2011 ◽  
Vol 57 (7) ◽  
pp. 1032-1041 ◽  
Author(s):  
Thomas Kroneis ◽  
Jochen B Geigl ◽  
Amin El-Heliebi ◽  
Martina Auer ◽  
Peter Ulz ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Whole Genome Amplification ◽  
Single Cells ◽  
Molecular Genetic ◽  
Target Cells ◽  
Whole Genome ◽  
Single Cell Level ◽  
Cell Level ◽  
Genome Amplification

BACKGROUND Analysis of chromosomal aberrations or single-gene disorders from rare fetal cells circulating in the blood of pregnant women requires verification of the cells' genomic identity. We have developed a method enabling multiple analyses at the single-cell level that combines verification of the genomic identity of microchimeric cells with molecular genetic and cytogenetic diagnosis. METHODS We used a model system of peripheral blood mononuclear cells spiked with a colon adenocarcinoma cell line and immunofluorescence staining for cytokeratin in combination with DNA staining with the nuclear dye TO-PRO-3 in a preliminary study to define candidate microchimeric (tumor) cells in Cytospin preparations. After laser microdissection, we performed low-volume on-chip isothermal whole-genome amplification (iWGA) of single and pooled cells. RESULTS DNA fingerprint analysis of iWGA aliquots permitted successful identification of all analyzed candidate microchimeric cell preparations (6 samples of pooled cells, 7 samples of single cells). Sequencing of 3 single-nucleotide polymorphisms was successful at the single-cell level for 20 of 32 allelic loci. Metaphase comparative genomic hybridization (mCGH) with iWGA products of single cells showed the gains and losses known to be present in the genomic DNA of the target cells. CONCLUSIONS This method may be instrumental in cell-based noninvasive prenatal diagnosis. Furthermore, the possibility to perform mCGH with amplified DNA from single cells offers a perspective for the analysis of nonmicrochimeric rare cells exhibiting genomic alterations, such as circulating tumor cells.

Centrifugal-Driven Droplet Generation Method with Minimal Waste for Single-Cell Whole Genome Amplification

2019 ◽  
Vol 91 (21) ◽  
pp. 13611-13619 ◽  
Author(s):  
Xingrui Li ◽  
Dongfeng Zhang ◽  
Weidong Ruan ◽  
Weizhi Liu ◽  
Kun Yin ◽  
...  
Keyword(s):  
Single Cell ◽  
Whole Genome Amplification ◽  
Droplet Generation ◽  
Whole Genome ◽  
Genome Amplification
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