Chimeric Gene Library Construction by a Simple and Highly Versatile Method Using Recombination-Dependent Exponential Amplification

2003 ◽  
Vol 19 (5) ◽  
pp. 1460-1467 ◽  
Author(s):  
A. Ikeuchi ◽  
Y. Kawarasaki ◽  
T. Shinbata ◽  
T. Yamane
Keyword(s):  
Chimeric Gene ◽  
Gene Library ◽  
Library Construction

scholarly journals Validation and standardization of DNA extraction and library construction methods for metagenomics-based human fecal microbiome measurements

Microbiome ◽  
2021 ◽  
Vol 9 (1) ◽  
Author(s):  
Dieter M. Tourlousse ◽  
Koji Narita ◽  
Takamasa Miura ◽  
Mitsuo Sakamoto ◽  
Akiko Ohashi ◽  
...  
Keyword(s):  
Dna Extraction ◽  
High Throughput Sequencing ◽  
Performance Metrics ◽  
Collaborative Study ◽  
Human Microbiome ◽  
Second Phase ◽  
Intermediate Precision ◽  
Fecal Microbiome ◽  
Library Construction ◽  
Wide Range

Abstract Background Validation and standardization of methodologies for microbial community measurements by high-throughput sequencing are needed to support human microbiome research and its industrialization. This study set out to establish standards-based solutions to improve the accuracy and reproducibility of metagenomics-based microbiome profiling of human fecal samples. Results In the first phase, we performed a head-to-head comparison of a wide range of protocols for DNA extraction and sequencing library construction using defined mock communities, to identify performant protocols and pinpoint sources of inaccuracy in quantification. In the second phase, we validated performant protocols with respect to their variability of measurement results within a single laboratory (that is, intermediate precision) as well as interlaboratory transferability and reproducibility through an industry-based collaborative study. We further ascertained the performance of our recommended protocols in the context of a community-wide interlaboratory study (that is, the MOSAIC Standards Challenge). Finally, we defined performance metrics to provide best practice guidance for improving measurement consistency across methods and laboratories. Conclusions The validated protocols and methodological guidance for DNA extraction and library construction provided in this study expand current best practices for metagenomic analyses of human fecal microbiota. Uptake of our protocols and guidelines will improve the accuracy and comparability of metagenomics-based studies of the human microbiome, thereby facilitating development and commercialization of human microbiome-based products.

cDNA library construction of two human Demodexspecies

2017 ◽  
Vol 62 (2) ◽  
Author(s):  
DongLing Niu ◽  
RuiLing Wang ◽  
YaE Zhao ◽  
Rui Yang ◽  
Li Hu ◽  
...  
Keyword(s):  
Cdna Library ◽  
Cdna Library Construction ◽  
Library Construction

AbstractThe research of

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