Semirational Approach for Ultrahigh Poly(3-hydroxybutyrate) Accumulation inEscherichia coliby Combining One-Step Library Construction and High-Throughput Screening

2016 ◽  
Vol 5 (11) ◽  
pp. 1308-1317 ◽  
Author(s):  
Teng Li ◽  
Jianwen Ye ◽  
Rui Shen ◽  
Yeqing Zong ◽  
Xuejin Zhao ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Library Construction ◽  
One Step

scholarly journals One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays

2016 ◽  
Vol 21 (10) ◽  
pp. 1112-1124 ◽  
Author(s):  
Sheng Dai ◽  
Rong Li ◽  
Yan Long ◽  
Steve Titus ◽  
Jinghua Zhao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Neural Stem Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Neuronal Cells ◽  
Drug Efficacy ◽  
Human Stem Cells ◽  
Induced Pluripotent Cells ◽  
One Step ◽  
Human Neuronal Cells

Human neuronal cells differentiated from induced pluripotent cells have emerged as a new model system for the study of disease pathophysiology and evaluation of drug efficacy. Differentiated neuronal cells are more similar in genetics and biological content to human brain cells than other animal disease models. However, culture of neuronal cells in assay plates requires a labor-intensive procedure of plate precoating, hampering its applications in high-throughput screening (HTS). We developed a simplified method with one-step seeding of neural stem cells in assay plates by supplementing the medium with a recombinant human vitronectin (VTN), thus avoiding plate precoating. Robust results were obtained from cell viability, calcium response, and neurite outgrowth assays using this new method. Our data demonstrate that this approach greatly simplifies high-throughput assays using neuronal cells differentiated from human stem cells for translational research.

scholarly journals Cas-CLIP: a method for customizing pooled CRISPR libraries

2018 ◽  
Author(s):  
Jiyeon Kweon ◽  
Da-eun Kim ◽  
An-Hee Jang ◽  
Yongsub Kim
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Target Genes ◽  
Biological Processes ◽  
Precise Method ◽  
Library Construction ◽  
Expensive Process ◽  
Screening Technology

ABSCTRACTAlthough pooled CRISPR libraries are widely used in high-throughput screening to study various biological processes, library construction for researcher’s own study is a time-consuming, labor-intensive, and expensive process. In this study, we develop a simple, scalable method, called Cas-CLIP, to customize conventional pooled CRISPR libraries using the CRISPR/Cas9 system. We show that conventional pooled CRISPR libraries can be modified by eliminating gRNAs that target positive genes, enabling the identification of unknown target genes in CRISPR screening. Cas-CLIP is a precise method for customizing conventional pooled CRISPR libraries and will broaden the scope of high-throughput screening technology.

ORIGINAL RESEARCH: A dedicated vector for efficient library construction and high throughput screening in the hyphal fungus Chrysosporium lucknowense

2007 ◽  
Vol 3 (1) ◽  
pp. 48-57 ◽  
Author(s):  
Jan C. Verdoes ◽  
Peter J. Punt ◽  
Richard Burlingame ◽  
Jeffrey Bartels ◽  
Reijer van Dijk ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Original Research ◽  
Library Construction

scholarly journals A High-Throughput Screening Method for Endophytic Bacteria With Antagonistic Activity Against Magnaporthe Oryzae in Rice (Oryza Sativa L.) Seeds

2021 ◽  
Author(s):  
Zhishan Wang ◽  
Xianyu Wu ◽  
Ni Li ◽  
Weiping Wang ◽  
Yang Liu
Keyword(s):  
High Throughput ◽  
Magnaporthe Oryzae ◽  
High Throughput Screening ◽  
Endophytic Bacteria ◽  
Oryza Sativa L ◽  
Screening Method ◽  
Antagonistic Activity ◽  
The Sustainable Development ◽  
One Step ◽  
Rice Seeds

Abstract Screening for microorganisms with antagonistic activity against pathogens in crops is of great significance for the preparation of microbial antagonists, the control of crop diseases, and the sustainable development of agriculture. Through the experiment, a rapid, efficient, and one-step high-throughput method for screening endophytic bacteria with antagonistic activity against Magnaporthe oryzae in rice seeds was successfully established, and the experimental results showed that the endophytic bacteria with antagonistic activity of Magnaporthe oryzae ACCC 36020 could be directly screened from six groups of experimental samples by this method. The establishment of this method can achieve simultaneous screening and purification, one-step high-throughput screening of antagonistic bacteria in rice seeds, and greatly save time.

scholarly journals Hi-TOM: a platform for high-throughput tracking of mutations induced by CRISPR/Cas systems

2017 ◽  
Author(s):  
Qing Liu ◽  
Chun Wang ◽  
Xiaozhen Jiao ◽  
Huawei Zhang ◽  
Lili Song ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
High Reliability ◽  
Mutation Screening ◽  
Screening Strategy ◽  
Library Construction ◽  
Genetic Modifications ◽  
Parameter Configuration ◽  
Next Generation Sequencing Ngs

AbstractThe CRISPR/Cas system has been extensively applied to make precise genetic modifications in various organisms. Despite its importance and widespread use, large-scale mutation screening remains time-consuming, labour-intensive and costly. Here, we describe a cheap, practicable and high-throughput screening strategy that allows parallel screening of 96 × N (N denotes the number of targets) genome-modified sites. The strategy simplified and streamlined the process of next-generation sequencing (NGS) library construction by fixing the bridge sequences and barcoding primers. We also developed Hi-TOM (available at http://www.hi-tom.net/hi-tom/), an online tool to track the mutations with precise percentage. Analysis of the samples from rice, hexaploid wheat and human cells reveals that the Hi-TOM tool has high reliability and sensitivity in tracking various mutations, especially complex chimeric mutations that frequently induced by genome editing. Hi-TOM does not require specially design of barcode primers, cumbersome parameter configuration or additional data analysis. Thus, the streamlined NGS library construction and comprehensive result output make Hi-TOM particularly suitable for high-throughput identification of all types of mutations induced by CRISPR/Cas systems.

Validation of Miniaturized One-Step Reverse Transcription qPCR Assays for High-Throughput Screening and Comparison to a Reporter Gene Methodology

2015 ◽  
Vol 13 (2) ◽  
pp. 94-101 ◽  
Author(s):  
Catherine Bardelle ◽  
Lisa McWilliams ◽  
Susan Mounfield ◽  
Mark Wigglesworth ◽  
Kirsty Rich
Keyword(s):  
Reporter Gene ◽  
High Throughput ◽  
Reverse Transcription ◽  
High Throughput Screening ◽  
One Step
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