DNA or GENE TRANSFER TECHNIQUES- PHYSICAL AND CHEMICAL (NOTES)
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PHYSICAL AND CHEMICAL METHODS OF DNA TRANSFER- MICROINJECTION, ELECTROPORATION, LIPOFECTION, PARTICLE GUN BOMBARDMENT, CALCIUM PHOSPHATE PRECIPITATION
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Site-directed mutagenesis is a molecular biology technique used to make specific changes to DNA sequences. It involves using a primer containing the desired mutation in a PCR reaction to introduce the mutation into the gene of interest. There are different approaches for site-directed mutagenesis using PCR, including using a mutated primer in normal PCR or a primer extension method. The technique is used for applications like protein engineering to study the impact of sequence changes or insert restriction sites. However, it can be difficult to replicate the mutated DNA and screening mutations requires sequencing.
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This document describes the blue-white screening technique for identifying recombinant bacteria. Blue-white screening relies on the activity of β-galactosidase, an enzyme in E. coli that cleaves lactose. Plasmid vectors used in cloning carry a fragment of the lacZ gene. When the plasmid inserts foreign DNA at the multiple cloning site, it disrupts lacZ and prevents complementation, so the bacteria cannot metabolize lactose and appear white on an indicator plate. If no DNA inserts or inserts elsewhere, complementation occurs and bacteria appear blue. The technique allows rapid identification of bacteria that took up recombinant plasmid.
Artificial Vectors
Artificial vectors such as bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), P1-derived artificial chromosomes (PACs), and human artificial chromosomes (HACs) can carry large DNA fragments for research purposes. BACs can hold up to 300kb of DNA and are used to sequence genomes. YACs can carry 500kb of DNA but are prone to rearrangement. HACs are separate microchromosomes that act as new chromosomes and can carry therapeutic genes for research including disease models.
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Reporter gene[2]
Reporter genes are genes that produce easily detectable and quantifiable proteins to track the expression of other genes. Common reporter genes include GFP, luciferase, CAT, and β-galactosidase. Reporter genes are used to study gene expression patterns, monitor plant transformation, and study regulatory elements. There are two main types of reporter genes - scorable markers, which produce a quantifiable phenotype, and selectable markers, which allow cells to survive under selective conditions using antibiotic resistance.
Blue white screening
This document describes the blue-white screening technique for identifying recombinant bacteria. Blue-white screening relies on the activity of β-galactosidase, an enzyme in E. coli that cleaves lactose. Plasmid vectors used in cloning carry a fragment of the lacZ gene. When the plasmid inserts foreign DNA at the multiple cloning site, it disrupts lacZ and prevents complementation, so the bacteria cannot metabolize lactose and appear white on an indicator plate. If no DNA inserts or inserts elsewhere, complementation occurs and bacteria appear blue. The technique allows rapid identification of bacteria that took up recombinant plasmid.
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Artificial vectors such as bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), P1-derived artificial chromosomes (PACs), and human artificial chromosomes (HACs) can carry large DNA fragments for research purposes. BACs can hold up to 300kb of DNA and are used to sequence genomes. YACs can carry 500kb of DNA but are prone to rearrangement. HACs are separate microchromosomes that act as new chromosomes and can carry therapeutic genes for research including disease models.
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Strain improvement and development involves manipulating microbial strains to enhance their metabolic capacities for biotechnology applications. Targets of improvement include rapid growth, genetic stability, non-toxicity, large cell size, ability to use cheaper substrates, increased productivity, and reduced cultivation costs. Methods for optimization include modifying environmental conditions, nutrition, mutagenesis, transduction, conjugation, transformation, and genetic engineering. Common industrial microorganisms are bacteria such as Bacillus subtilis and yeasts such as Saccharomyces cerevisiae.
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Transformation and transfection
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Site-directed mutagenesis is a technique used to introduce specific changes to the DNA sequence of a gene by altering the nucleotide sequence. It allows researchers to study the impact of mutations by changing individual bases, deleting bases, or inserting new bases. There are different methods of site-directed mutagenesis including oligonucleotide-based methods and PCR-based methods. Site-directed mutagenesis has applications in research, production of desired proteins, and development of engineered proteins for commercial uses like detergents.
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This document discusses methods for improving microbial strains through genetic modification to increase production of commercially valuable metabolites. There are two main methods - mutation and genetic recombination. Mutation involves inducing changes in a microorganism's DNA through mutagenic agents like UV light or chemicals to generate mutants with desirable traits like higher yields, antibiotic resistance, or growth on low-cost substrates. Genetic recombination combines genetic material from two strains, such as combining a high yielding mutant with a wild type or another mutant, to further increase fermentation yields.
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1. Cosmid vectors are cloning vectors derived from bacteriophages that can contain up to 44 kilobase pairs of foreign DNA. They are commonly used to clone large fragments of genomic DNA in E. coli.
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This document discusses tower fermenters, which are elongated fermentation vessels with a height to width aspect ratio of 6:1 or more that allow for the unidirectional flow of gases. There are several types of tower fermenters including bubble columns, vertical tower beer fermenters, and multistage fermenter systems. Tower fermenters have been used for the production of products such as citric acid, tetracycline, beer, and to cultivate organisms like yeast and E. coli. They provide a simple design for aerobic fermentation of cells and enzymes.
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the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
Shallowest Oil Discovery of Turkiye.pptx
The Petroleum System of the Çukurova Field - the Shallowest Oil Discovery of Türkiye, Adana
SAR of Medicinal Chemistry 1st by dk.pdf
In this presentation include the prototype drug SAR on thus or with their examples .
Syllabus of Second Year B. Pharmacy
2019 PATTERN.
在线办理(salfor毕业证书)索尔福德大学毕业证毕业完成信一模一样
学校原件一模一样【微信：741003700 】《(salfor毕业证书)索尔福德大学毕业证》【微信：741003700 】学位证，留信认证（真实可查，永久存档）原件一模一样纸张工艺/offer、雅思、外壳等材料/诚信可靠,可直接看成品样本，帮您解决无法毕业带来的各种难题！外壳，原版制作，诚信可靠，可直接看成品样本。行业标杆！精益求精，诚心合作，真诚制作！多年品质 ,按需精细制作，24小时接单,全套进口原装设备。十五年致力于帮助留学生解决难题，包您满意。
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◇申请留学生创业基金
留信网认证的作用:
1:该专业认证可证明留学生真实身份
2:同时对留学生所学专业登记给予评定
3:国家专业人才认证中心颁发入库证书
4:这个认证书并且可以归档倒地方
5:凡事获得留信网入网的信息将会逐步更新到个人身份内，将在公安局网内查询个人身份证信息后，同步读取人才网入库信息
6:个人职称评审加20分
7:个人信誉贷款加10分
8:在国家人才网主办的国家网络招聘大会中纳入资料，供国家高端企业选择人才
The debris of the ‘last major merger’ is dynamically young
The Milky Way’s (MW) inner stellar halo contains an [Fe/H]-rich component with highly eccentric orbits, often referred to as the
‘last major merger.’ Hypotheses for the origin of this component include Gaia-Sausage/Enceladus (GSE), where the progenitor
collided with the MW proto-disc 8–11 Gyr ago, and the Virgo Radial Merger (VRM), where the progenitor collided with the
MW disc within the last 3 Gyr. These two scenarios make different predictions about observable structure in local phase space,
because the morphology of debris depends on how long it has had to phase mix. The recently identified phase-space folds in Gaia
DR3 have positive caustic velocities, making them fundamentally different than the phase-mixed chevrons found in simulations
at late times. Roughly 20 per cent of the stars in the prograde local stellar halo are associated with the observed caustics. Based
on a simple phase-mixing model, the observed number of caustics are consistent with a merger that occurred 1–2 Gyr ago.
We also compare the observed phase-space distribution to FIRE-2 Latte simulations of GSE-like mergers, using a quantitative
measurement of phase mixing (2D causticality). The observed local phase-space distribution best matches the simulated data
1–2 Gyr after collision, and certainly not later than 3 Gyr. This is further evidence that the progenitor of the ‘last major merger’
did not collide with the MW proto-disc at early times, as is thought for the GSE, but instead collided with the MW disc within
the last few Gyr, consistent with the body of work surrounding the VRM.
Compexometric titration/Chelatorphy titration/chelating titration
Classification
Metal ion ion indicators
Masking and demasking reagents
Estimation of Magnisium sulphate
Calcium gluconate
Complexometric Titration/ chelatometry titration/chelating titration, introduction, Types-
1.Direct Titration
2.Back Titration
3.Replacement Titration
4.Indirect Titration
Masking agent, Demasking agents
formation of complex
comparition between masking and demasking agents,
Indicators/Metal ion indicators/ Metallochromic indicators/pM indicators,
Visual Technique,PM indicators (metallochromic), Indicators of pH, Redox Indicators
Instrumental Techniques-Photometry
Potentiometry
Miscellaneous methods.
Complex titration with EDTA.
Equivariant neural networks and representation theory
Or: Beyond linear.
Abstract: Equivariant neural networks are neural networks that incorporate symmetries. The nonlinear activation functions in these networks result in interesting nonlinear equivariant maps between simple representations, and motivate the key player of this talk: piecewise linear representation theory.
Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
Corrected slides: dtubbenhauer.com/talks.html
The binding of cosmological structures by massless topological defects
Assuming spherical symmetry and weak field, it is shown that if one solves the Poisson equation or the Einstein field
equations sourced by a topological defect, i.e. a singularity of a very specific form, the result is a localized gravitational
field capable of driving flat rotation (i.e. Keplerian circular orbits at a constant speed for all radii) of test masses on a thin
spherical shell without any underlying mass. Moreover, a large-scale structure which exploits this solution by assembling
concentrically a number of such topological defects can establish a flat stellar or galactic rotation curve, and can also deflect
light in the same manner as an equipotential (isothermal) sphere. Thus, the need for dark matter or modified gravity theory is
mitigated, at least in part.
The cost of acquiring information by natural selection
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
Randomised Optimisation Algorithms in DAPHNE
Slides from talk:
Aleš Zamuda: Randomised Optimisation Algorithms in DAPHNE .
Austrian-Slovenian HPC Meeting 2024 – ASHPC24, Seeblickhotel Grundlsee in Austria, 10–13 June 2024
https://ashpc.eu/
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...Advanced-Concepts-Team
Presentation in the Science Coffee of the Advanced Concepts Team of the European Space Agency on the 07.06.2024.
Speaker: Diego Blas (IFAE/ICREA)
Title: Gravitational wave detection with orbital motion of Moon and artificial
Abstract:
In this talk I will describe some recent ideas to find gravitational waves from supermassive black holes or of primordial origin by studying their secular effect on the orbital motion of the Moon or satellites that are laser ranged.Recently uploaded (20)
Immersive Learning That Works: Research Grounding and Paths Forward
Immersive Learning That Works: Research Grounding and Paths Forward
ESR spectroscopy in liquid food and beverages.pptx
ESR spectroscopy in liquid food and beverages.pptx
(June 12, 2024) Webinar: Development of PET theranostics targeting the molecu...
(June 12, 2024) Webinar: Development of PET theranostics targeting the molecu...
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
THEMATIC APPERCEPTION TEST(TAT) cognitive abilities, creativity, and critic...
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...
aziz sancar nobel prize winner: from mardin to nobel
aziz sancar nobel prize winner: from mardin to nobel
GBSN - Biochemistry (Unit 6) Chemistry of Proteins
GBSN - Biochemistry (Unit 6) Chemistry of Proteins
The debris of the ‘last major merger’ is dynamically young
The debris of the ‘last major merger’ is dynamically young
Compexometric titration/Chelatorphy titration/chelating titration
Compexometric titration/Chelatorphy titration/chelating titration
Equivariant neural networks and representation theory
Equivariant neural networks and representation theory
The binding of cosmological structures by massless topological defects
The binding of cosmological structures by massless topological defects
The cost of acquiring information by natural selection
The cost of acquiring information by natural selection
Basics of crystallography, crystal systems, classes and different forms
Basics of crystallography, crystal systems, classes and different forms
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...
ESA/ACT Science Coffee: Diego Blas - Gravitational wave detection with orbita...