Evidence map›Paper›PMID 30461059›Full record

ArticleJournal of clinical laboratory analysis2019

A low cost and input tailing method of quality control on multiple annealing, and looping-based amplification cycles-based whole-genome amplification products.

Changyue Chen, Jing Li, JueFeng Wan, Yuan Lu, Zhen Zhang, ZengHui Xu

Abstract read
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Article in Journal of clinical laboratory analysis, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

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4citing papers in PubMed
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1 · What the graph read from it

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3 · Its place in the literature

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4 citing papers in PubMed.

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5 · Who and what money

Authors and funding

6 authors.

Changyue ChenDepartment of Medical Research, Shanghai MajorMed Diagnostics Company, Shanghai, China.ORCID http://orcid.org/0000-0002-3534-6701
Jing LiDepartment of Medical Research, Shanghai MajorMed Diagnostics Company, Shanghai, China.
JueFeng WanDepartment of Radiation Oncology, Fudan University Shanghai Cancer Center, Shanghai, China.
Yuan LuDepartment of Medical Research, Shanghai MajorMed Diagnostics Company, Shanghai, China.
Zhen ZhangDepartment of Radiation Oncology, Fudan University Shanghai Cancer Center, Shanghai, China.
ZengHui XuLaboratory of Gene and Viral Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University of Chinese PLA, Shanghai, China.

Funding

National Natural Science Foundation of China 81372432
6 · The paper itself

Abstract

backgroundSingle-cell whole-genome sequencing provides novel insights into the nature of genetic heterogeneity in normal and diseased cells. However, amplification of formalin-fixed tissues with low cell numbers is still problematic and multiple annealing, and looping-based amplification cycles (MALBAC) is a commonly used whole-genome amplification (WGA) method with low cell numbers.

methodsWe developed a low-input tailing method to evaluate the MALBAC-based WGA from sub-nanogram or less quantities of input DNA. The tailing method uses 2100 BioAnalyzer to evaluate the size distribution of MALBAC products, and comparing the tailing with 10380 bp.

resultsCompared with a 22 loci qPCR panel, the tailing method provided a similar WGA evaluation efficiency in 13 samples on one set of study, with lower input, cheaper cost, shorter manual time, and a clear filtering cut off. Later, we demonstrated a strong correlation between tailing size and coverage breadth in another 29 samples on two sets of assays. As a result, the tailing method showed that it could predict whether a sequence breadth achieved 70% or not with 100% accuracy on these three sets of assays. Although further studies are needed, this tailing method is expected to be used as an excellent tool to select high-quality WGA products before library construction.

conclusionsOur tailing method can provide a new WGA quality test to evaluate the WGA efficiency with 100% accuracy (42/42). Compared with qPCR panel, our tailing method needs lower input, cheaper cost, shorter manual time, a clear filtering cut off, and extendable high throughput as well as the same sensitivity.

Indexed as

GenomicsNucleic Acid Amplification TechniquesGenome, HumanHumansQuality Control

Identifiers

PMID30461059
PMCPMC6818575

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.