scholarly journals Antha-guided Automation of Darwin Assembly for the Construction of Bespoke Gene Libraries

2019 ◽  
Author(s):  
P. Handal Marquez ◽  
M. Koch ◽  
D. Kestemont ◽  
S. Arangundy-Franklin ◽  
V. B. Pinheiro
Keyword(s):  
Saturation Mutagenesis ◽  
Liquid Handling ◽  
Assembly Method ◽  
Amplicon Length ◽  
Hands On ◽  
Experimental Parameters ◽  
Gene Libraries ◽  
Fast Reconstruction ◽  
Protein Libraries

AbstractProtein engineering through directed evolution facilitates the screening and characterization of protein libraries. Efficient and effective methods for multiple site-saturation mutagenesis, such as Darwin Assembly, can accelerate the sampling of relevant sequence space and the identification of variants with desired functionalities. Here, we present the automation of the Darwin Assembly method, using a Gilson PIPETMAX™ liquid handling platform under the control of the Antha software platform, which resulted in the accelerated construction of complex, multiplexed gene libraries with minimal hands-on time and error-free, while maintaining flexibility over experimental parameters through a graphical user interface rather than requiring user-driven library-dependent programming of the liquid handling platform. We also present an approach for barcoding libraries that overcomes amplicon length limitations in next generation sequencing and enables fast reconstruction of library reads.

Preparation and Characterization of Polyaniline Nanotubes Doped with Different Acid

2013 ◽  
Vol 401-403 ◽  
pp. 663-666
Author(s):  
Xue Lian Bai ◽  
Jian Ting Mei ◽  
Zhong Guo Mu ◽  
Yun Bai
Keyword(s):  
Acetic Acid ◽  
Self Assembly ◽  
Ammonium Persulfate ◽  
Tetraacetic Acid ◽  
X Ray Diffraction ◽  
X Ray ◽  
Structure And Property ◽  
Assembly Method ◽  
Polyaniline Nanotubes

Polyaniline (PANI) nanotubes were synthesized separately using amino acetic acid (AA), ethylenediamine tetraacetic acid (EDTA), oxalic acid (OA) as dopant and ammonium persulfate (APS) as oxidant by a self-assembly method. SEM, TEM,FTIR and X-ray diffraction (XRD) and applying the 4 probes method characterized the morphology, structure and property of the product. It was found that nanotubes morphology were synthesized when the [Aci/[A ratio is 1:2.The room template conductivity of the products were studied.

Sensitivity analysis of BISON model for characterization of impact of experimental parameters on hydrogen migration and redistribution in zirconium-based alloys

2021 ◽  
Vol 550 ◽  
pp. 152941
Author(s):  
Seok Bin Seo ◽  
Edward Matthew Duchnowski ◽  
Miles O'Neal ◽  
Arthur T. Motta ◽  
Florian Passelaigue ◽  
...  
Keyword(s):  
Sensitivity Analysis ◽  
Hydrogen Migration ◽  
Experimental Parameters

Experimental Parameters Used to Study Pesticide Degradation in Soil

1996 ◽  
Vol 10 (1) ◽  
pp. 169-173 ◽  
Author(s):  
Michael R. Blumhorst
Keyword(s):  
Small Scale ◽  
Test Substance ◽  
Pesticide Degradation ◽  
Soil System ◽  
Experimental Parameters ◽  
Effective Cost ◽  
Cost Efficient ◽  
Efficient Mechanisms ◽  
Degradation In Soil

Characterization of pesticide degradation in soil is an important component in determining the environmental impact of agriculturally-applied pesticides. Several techniques currently are being used to generate these data, but small-scale laboratory studies remain one of the most effective, cost-efficient mechanisms of evaluating pesticide behavior in soil. With small-scale studies, many different environmental factors can be incorporated into the experimental design, and with the use of14C-labeled material, these studies (often referred to as soil degradation or soil metabolism studies) provide information on test substance persistence, degradation, volatilization, and mineralization. Care must be exercised, however, in selecting the experimental parameters to be used because of the potential adverse or artificial effects on the soil system.

Characterization of T-DNA Insertion Sites in Arabidopsis thaliana and the Implications for Saturation Mutagenesis

2002 ◽  
Vol 6 (2) ◽  
pp. 163-174 ◽  
Author(s):  
Patrick J. Krysan ◽  
Jeffery C. Young ◽  
Peter J. Jester ◽  
Sean Monson ◽  
Greg Copenhaver ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Saturation Mutagenesis ◽  
Dna Insertion ◽  
Insertion Sites

Enzymatic Thioxyloside Synthesis: Characterization of Thioglycoligase Variants Identified from A Site-Saturation Mutagenesis Library ofBacillus CirculansXylanase

ChemBioChem ◽  
2010 ◽  
Vol 11 (4) ◽  
pp. 533-538 ◽  
Author(s):  
Zachary Armstrong ◽  
Stephan Reitinger ◽  
Terrence Kantner ◽  
Stephen G. Withers
Keyword(s):  
Saturation Mutagenesis ◽  
Site Saturation ◽  
A Site

Molecular cloning and characterization of mutant and wild-type human beta-actin genes

1984 ◽  
Vol 4 (10) ◽  
pp. 1961-1969
Author(s):  
J Leavitt ◽  
P Gunning ◽  
P Porreca ◽  
S Y Ng ◽  
C S Lin ◽  
...  
Keyword(s):  
Homologous Recombination ◽  
Human Fibroblasts ◽  
Actin Gene ◽  
Actin Genes ◽  
Beta Actin ◽  
Gene Libraries ◽  
Actin Mrna ◽  
Dna Fragment ◽  
Specific Sequences

There are more than 20 beta-actin-specific sequences in the human genome, many of which are pseudogenes. To facilitate the isolation of potentially functional beta-actin genes, we used the new method of B. Seed (Nucleic Acids Res. 11:2427-2446, 1983) for selecting genomic clones by homologous recombination. A derivative of the pi VX miniplasmid, pi AN7 beta 1, was constructed by insertion of the 600-base-pair 3' untranslated region of the beta-actin mRNA expressed in human fibroblasts. Five clones containing beta-actin sequences were selected from an amplified human fetal gene library by homologous recombination between library phage and the miniplasmid. One of these clones contained a complete beta-actin gene with a coding sequence identical to that determined for the mRNA of human fibroblasts. A DNA fragment consisting of mostly intervening sequences from this gene was then used to identify 13 independent recombinant copies of the analogous gene from two specially constructed gene libraries, each containing one of the two types of mutant beta-actin genes found in a line of neoplastic human fibroblasts. The amino acid and nucleotide sequences encoded by the unmutated gene predict that a guanine-to-adenine transition is responsible for the glycine-to-aspartic acid mutation at codon 244 and would also result in the loss of a HaeIII site. Detection of this HaeIII polymorphism among the fibroblast-derived clones verified the identity of the beta-actin gene expressed in human fibroblasts.

Preparation and characterization of poly(styrenemethacrylic acid)/MCM-41 core/shell nanocomposite microspheres

e-Polymers ◽  
2009 ◽  
Vol 9 (1) ◽  
Author(s):  
Yansheng Zhao ◽  
Xingji Ma ◽  
Yongmei Liu ◽  
Guangwei Yuan ◽  
Meijuan Guo ◽  
...  
Keyword(s):  
Self Assembly ◽  
Cetyltrimethylammonium Bromide ◽  
Average Diameter ◽  
Molar Ratio ◽  
Core Shell ◽  
Acidic Media ◽  
Assembly Method ◽  
20 Nm ◽  
Mcm 41

AbstractIn acidic media, poly(styrene-methacrylic acid)/MCM-41 [P(St- MAA)/MCM-41] core/shell microspheres were synthesized using monodisperse P(St-MAA) particles contained in soap-free emulsion and cetyltrimethylammonium bromide as co-templates by adsorption self-assembly method. The effects of P(St- MAA) composition on shell structure of the core/shell microspheres were investigated. The morphology and composition of P(St-MAA)/MCM-41 microspheres were characterized by TEM, XRD and FTIR. The results show that the ordering degree of MCM-41 shells increased as the molar ratio of MAA to St increased. When n(MAA)/n(St) is 0.2, the average diameter and the shell thickness of nanocomposite microspheres are about 170 nm and 20 nm, respectively.

Quality Control Procedures in Herbicide Field Dissipation Studies

1997 ◽  
Vol 11 (4) ◽  
pp. 832-837 ◽  
Author(s):  
Michael R. Blumhorst ◽  
Thomas C. Mueller
Keyword(s):  
Quality Control ◽  
Environmental Impact ◽  
Experimental Approach ◽  
Recent Literature ◽  
Research Area ◽  
Field Conditions ◽  
Experimental Parameters ◽  
Control Procedures ◽  
Areas Of Concern

Characterization of herbicide dissipation under field conditions is an important component in determining the environmental impact of herbicides, and much information is available on this subject. However, this research area could be improved by standardizing and implementing several quality control procedures. The purpose of this review is to evaluate the recent literature on herbicide dissipation published in journals of the Weed Science Society of America. This review is not comprehensive with respect to all experimental parameters associated with field dissipation studies, but it is aimed at addressing specific areas of concern in the experimental approach to this research.

Sign in / Sign up

Export Citation Format

Share Document