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We narrowed to 608 results for: Sis

Showing: 1 - 40 of 608 results

  1. Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

    Type
    Blog Post
    Published
    April 12, 2018, 1:02 p.m.
    ... two global stock centers for Arabidopsis thaliana (Arabidopsis). Our mission is to collect, preserve,...in 60 countries each year. Introducing Arabidopsis Arabidopsis is an important model system for research...and you have yourself a plant publicity crisis. The Arabidopsis Biological Resource Center (ABRC) set out...Translating Research on Arabidopsis Into a Network of Educational Resources). Arabidopsis is used in research... the Education & Outreach Specialist for the Arabidopsis Biological Resource Center and the Center for...Ohio State University. Established in 1991, the Arabidopsis Biological Resource Center (ABRC) is one of two...distribute seeds, DNA and other resources for Arabidopsis and related species. Located at The Ohio State...

  2. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    Published
    Feb. 21, 2019, 1:15 p.m.
    ...spectinomycin resistant phenotype in bacteria. While there are many known spectinomycin resistance mutations...genomic site for mutagenesis, a feature which they used to generate bacteria resistant to both spectinomycin...directed mutagenesis to generate specific mutations in your plasmids Use REPLACER Mutagenesis to mutate...Mutagenesis is a tool that both evolution and molecular biologists use to tinker with DNA. Making changes...region. EvolvR, a CRISPR-Cas9 based targeted mutagenesis method developed by the Dueber Lab at Berkeley...potential applications. Challenges of existing mutagenesis methods Several continuous diversification methods...error prone polymerase I (PolI3M) is capable of mutagenesis in E. Coli, but this approach requires special...

  3. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ..., a graduate student at Stanford University. Arabidopsis is a fantastic model organism for many reasons... are many motivations to generate transgenic Arabidopsis, from studying transcriptional and translational... plants, to complementing mutant phenotypes. Arabidopsis is amenable to the floral drip or dip transformation...transformation protocol, as well as protocols for growing Arabidopsis are also available (Weigel and Glazebrook, 2002...vector is the first step in generating transgenic Arabidopsis. There are a bunch of great options, but here... pool you will be selecting from later. The Arabidopsis transformation process The process itself is ... the chances of tDNA integration. Selecting Arabidopsis transformants Once your seeds from your T0 plants...

  4. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ... the mutagenesis reaction. Alternative approaches to site directed mutagenesis Point-mutagenesis is fairly...constructs), in functional analysis of proteins (to perform alanine scanning mutagenesis or targeted substitution...should be resistant to Cas9 cleavage. If a plasmid contains the template, site-directed mutagenesis can be...resulting construct resistant to Cas9 induced cleavage. Summary of Site Directed Mutagenesis In brief, point-mutations...mutation of interest. Gel electrophoresis diagrams of the anticipated restriction analysis results (left) and...Laursen from Cornell University. Site directed mutagenesis is a highly versatile technique that can be used...substitution of key residues), and in SNP analysis (to introduce naturally occuring SNPs in a plasmid context...

  5. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    Published
    Jan. 30, 2014, 3:29 p.m.
    ...protein synthesis Bactericidal 25-100 ug/mL Ampicillin beta-lactam Inhibits cell wall synthesis Bactericidal...Resistance to antibiotics is a widely used tool in molecular biology, yet scientists rarely stop to think...to maintain the plasmid. Adding an antibiotic resistance gene to the plasmid solves both problems at once...pressure to keep your plasmid. Viva la (bacterial) resistance!  What are antibiotics? Antibiotics are generally...Binds 30S ribosomal subunit; interrupts protein synthesis Bactericidal 7.5-50 ug/mL Streptomycin aminoglycoside...Carbenicillin beta-lactam Inhibits cell wall synthesis Bactericidal 100 ug/mL Bleomycin glycopeptide...Binds 30S ribosomal subunit; inhibits protein synthesis (elongation step) Bacteriostatic 10 ug/mL ...

  6. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    Published
    Nov. 10, 2016, 3:30 p.m.
    ...Microbiology Pages What is antibiotic resistance? Antibiotic resistance is defined as microbial survival despite...generally organisms with multi-drug resistance are resistant to three or more different classes, or types...Learn How Antibiotic Resistance Genes Are Used in the Lab How does antibiotic resistance develop? There are...in which bacteria can become resistant to antibacterial drugs. Resistance can be intrinsic, meaning a ...means the bacterium is not naturally resistant but has become resistant somehow overtime. One fantastic example... example of intrinsic resistance would be Mycoplasma species’ resistance to beta-lactam antibiotics, like...Mycoplasma are intrinsically resistant to this class of antibiotic. Acquired resistance can be the result of ...

  7. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...beta-lactam that inhibits cell wall synthesis. The AmpR (amp resistance) gene produces beta-lactamase to ...processes (e.g., protein synthesis), while amp only affects the cell wall synthesis step during cell division...aminoglycoside which inhibits protein synthesis. The kan resistance gene, NPTII, facilitates phosphorylation...cells, are already resistant to this antibiotic due to stable integration of resistance genes in their genomes...damage. Choosing an antibiotic resistance Choosing the best antibiotic resistance ultimately comes down to ...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...interest. But does it matter which antibiotic resistance gene you select? In this blog, we’ll break down...

  8. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...this insertion size, primer synthesis is more expensive than gBlock synthesis, and multifragment assembly...REPLACR-mutagenesis could save busy scientists a lot of time. Let us know how REPLACR-mutagenesis works ...Comparison of REPLACR-mutagenesis with other common methods for site directed mutagenesis   References 1. ...al. (2016). “REPLACR-mutagenesis: a one-step method for site-directed mutagenesis by recombineering.” ...Site-directed mutagenesis (SDM) is one of the key tools researchers use to prove causation in molecular...vitro assembly methods. A new method, REPLACR-mutagenesis, harnesses the power of bacterial recombineering... Plasmids After PCR). Current site directed mutagenesis methods Current SDM methods rely on in vitro ...

  9. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ...interest, C-terminal tags, plant terminators and resistance cassettes for selection of transgenic plants....Lohmann said they’ve only tested GreenGate in Arabidopsis, it should in principle work in any plant species...Versatile and Efficient Cloning System for Plant Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013)...

  10. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    Published
    May 21, 2019, 12:10 p.m.
    ...Apoptosis or “programmed cell death" plays a pivotal role in an array of biological processes including...development, the immune system, and cell turnover. Apoptosis is a highly controlled process that is triggered...fragmentation and death (Elmore, 2007). To study apoptosis in vivo, scientists have used genetically encoded...fluorescent executioner caspase reporters. Several apoptosis reporters have been developed, such as the Forster...Zhang et al., 2019). Thus to effectively study apoptosis in vivo scientists need a protease reporter that...brightness. The Shu Lab from UCSF designed an improved apoptosis reporter, ZipGFP, which utilized the self-assembling... better study the spatio-temporal dynamics of apoptosis in living animals. To do this they modified a ...

  11. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...effects. One-pot saturation mutagenesis The one-pot saturation mutagenesis technique described in Wrenbeck... as the gene or use custom gene synthesis or site directed mutagenesis to remove the BbvCI site(s). ...saturation mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How .... “Random Mutagenesis by Error-Prone PCR”. Methods in Molecular Biology In Vitro Mutagenesis Protocols...the Whitehead lab developed One pot saturation mutagenesis as a quick and easy technique that can be used...library generation prior to one pot saturation mutagenesis Several methods can be used to create mutational... shortcomings compared to one pot saturation mutagenesis. Error-prone PCR: This method amplifies the...

  12. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...take a look at our live examples: experiment or analysis. In the rest of this article, we'll explain the...batch guide design, batch primer design or batch analysis. We wanted to relieve scientists of the burden... by Luca Pinello and Kendell Clement for batch analysis because of its ease-of-use and comprehensive reports...publicly-available for high-throughput design and analysis of CRISPR experiments. It has certainly boosted...

  13. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    Published
    June 26, 2018, 1:33 p.m.
    ...research on diseases of glycosylation, arginine mutagenesis, and neural development. Paula Videira: Congenital...disorders of glycosylation Paula Videira is an Assistant Professor at the Faculty of Sciences of NOVA University...functional activity of VUSs will enable more accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith...

  15. Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids

    Type
    Blog Post
    Published
    Nov. 20, 2018, 8:17 p.m.
    ... more long-term solution is the use of unique persistent identifiers (PIDs), a long-lasting way to identify...documents, files, or physical reagents. What are persistent identifiers? Perhaps the most commonly known ...original article online. It is also designed to be persistent and exist online indefinitely. PIDs historically...plasmids! In an effort to support standardized persistent identification of research reagents, several ...this post and sharing her technical expertise in persistent identifiers. Additional Resources on the Addgene...

  16. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    Published
    Nov. 27, 2018, 2 p.m.
    ...Mycobacterium tuberculosis resistance to pyrazinamide (PZA) (a drug commonly used to treat tuberculosis) and in...projects.   Understanding drug resistance mechanisms of Mycobacterium tuberculosis Dr. Patricia Sheen, Universidad... accurate detection of PZA-resistance is necessary to improve tuberculosis care and control. In her current...methods and improved therapies for PZA-resistant Mycobacteria tuberculosis. Investigation of multispecies biofilm...Sheen aims to identify new genes related to PZA-resistance by using different approaches, including CRISPR...experience in the field of bacterial and fungal pathogenesis. Her research focuses on a collection of pathogenic...

  17. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ...Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Cell. 2014...transcriptional activation of endogenous genes and it consists of three components: 1. A nucleolytically inactive... a synthetic transcription activation complex consisting of multiple distinct effector domains that work...genes that, when turned on, make melanoma cells resistant to treatment with a BRAF inhibitor. (The test ...-mediated screens exhibited a high degree of consistency and validation with >80% effectively enriched...

  18. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...only measures infectious viral particles. One of the si mplest methods of measuring functional titer uses...components such as LTRs, gag, WPRE, antibiotic resistance-genes, or the transgene itself. Many researchers...Another popular approach utilizes the antibiotic resistance genes carried on transfer plasmids. Target cells... one copy integrates, selecting by antibiotic-resistance or single cell sorting based on expression of...

  21. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...Brown W, Bardhan A, Delaney M, Ilk AS, Rauen RR, Kahn SI, Tsang M, Deiters A (2020) Spatiotemporal Control...protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9-CIBN, where CIBN is the N-terminal...(Kawano et al., 2015). Nicknamed paCas9-1 and consisting of the fusion proteins N713-pMag and nMagHigh1...Michael Lin, who developed a chimeric protein consisting of Cas9 and a photo-dissociable dimeric green...

  22. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...without a resistance gene, allowing resistant bacteria to multiply, spreading their resistance genes throughout...The crisis of antibiotic resistance is upon us, and the world is unprepared. Each year in the United ...antibiotic resistant bacteria. Even when researchers develop new antibiotics, the onset of resistance is swift...antibiotic resistant bacteria with a weapon that they’ll have less potential to become resistant to? CRISPR...antibiotic resistance Most antibiotics target one of three essential processes: cell wall synthesis, DNA replication...or antibiotic resistant bacteria, and inappropriate antibiotic usage selects for resistant bacteria, allowing...propagate their resistance genes. Our society is hoping to combat antibiotic resistance using multiple ...

  23. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...example, we examined ICE analysis of Sanger sequencing data alongside the analysis of NGS-based amplicon ...scalability, the ICE software has two analysis formats: sample by sample analysis, which can compare up to five...synthego.com. Overview of ICE editing analysis Once the analysis is complete, a new screen appears with...also download the entire analysis as a ZIP file by clicking the Download Analysis Data button on the bottom...bottom right of the analysis screen. ICE analysis details Contributions   The Contributions tab shows the...tool was initially created to support the CRISPR analysis needs of Synthego’s scientists, and it’s now free...sequencing data and found that the accuracy of ICE analysis results was highly comparable (within r2=0.96)...

  24. Deep Dive: qPCR

    Type
    Blog Post
    Published
    Sept. 29, 2022, 1:15 p.m.
    ...   Statistical analysis  Getting your RQ value isn’t the end of your analysis process. You’ll still...different statistical analysis methods that pair well with slightly modified CT analysis, so pick one and ...chemistries, primer optimization, controls, and analysis.  As a rule of thumb, you can assume that qPCR...chemistry  The two most common qPCR chemistries are hydrolysis probes (which you probably know as TaqMan) and...multiplexed Single amplicon validation technique Hydrolysis Probes (Taqman) Low; can be reduced with a ... Yes, one Yes Yes Agarose gel or uMelt analysis dsDNA dye (Sybr)  High No No No ... Yes, two Yes Yes Agarose gel or uMelt analysis Hairpin probes Low Yes, one Yes Yes...

  25. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...cancer development, metastasis, and drug resistance, Bhang says “the [drug] resistance story panned out ...cells that represent the preexisting resistant subpopulation 2. Resistant cells among the separate replicates...found that similar barcodes consistently made up the majority of the resistant populations. In fact, 88%...apply [it to].” Using ClonTracer to study drug resistance Although Bhang and co-workers started using the...determine whether or not cancer cell populations resistant to drug treatments arise during treatment or are... outcomes (see schematic depiction below): 1. Resistant cells among the separate replicates would contain...abundance before and after a treatment. The library consists of a pool of lentiviral vectors with roughly 73...

  26. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ...is as a biomedical model organism.  Arabidopsis thaliana Arabidopsis thaliana is one of, if not the most...manipulation and mutagenesis techniques, are available to the scientific community.  The Arabidopsis genome project...beginning of widespread Arabidopsis genetics research development. Arabidopsis thaliana was fully sequenced...for use in CRISPR and Tol2 transposase induced transgenesis.  Xenopus Xenopus, or frogs, are one of the ...understand human disease alleles during and after embryogenesis. Addgene has over 600 plasmids that contain ...characterized genome, and widespread utilization, Arabidopsis has made significant contributions to understanding...understanding plant and crop science alike.  Arabidopsis has many of the traits that make an ideal model organism...

  27. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ...have a plan to ensure it persists in both contexts. For a vector to persist in... You need... Bacteria... an antibiotic resistance gene to allow for antibiotic selection, but these resistance systems don’t always... antibiotics also block protein synthesis, and the neomycin resistance gene protects against any of them...single lecture slide. That shared language is the basis of shuttle vectors, which can function in unrelated...like penicillin and ampicillin inhibit cell wall synthesis. Eukaryotes, with no cell walls, can grow alongside...eukaryotes. For example, puromycin inhibits protein synthesis across kingdoms by targeting a shared aspect of... of ribosomal function. A puromycin resistance gene on your vector can protect both plasmid-carrying bacteria...

  28. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ... designed to insert a resistance gene. The vector contains a neomycin resistance gene (NeoR) flanked by...insert and later remove a resistance gene. In this construct, the neomycin resistance gene is floxed so that...into the vector on either side of an antibiotic resistance gene. To avoid selecting for cells in which the...perform a positive selection for the antibiotic resistance gene and then a counter selection for the negative...removed from the chromosome and and replaced by the resistance gene.The gene is thus disrupted, or knocked out...occurred. Neomycin, puromycin, and hygromycin resistance genes are commonly used for positive selection...positive selection marker. Since an antibiotic resistance gene can affect a cell's phenotype, researchers...

  29. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    Published
    June 2, 2016, 2:30 p.m.
    ...well as you perform analysis after analysis after analysis… Treat every analysis like a wet lab experiment... my thesis project a few years ago when my advisor and I decided to do a massive RNA-seq analysis. I had...you to perform this analysis. Having a clearly stated, specific goal for each analysis will help you locate...Before you begin, briefly state the goal of the analysis, including background on other analyses that have...approach that you will take to help you plan your analysis. There is no need to go into a lot of detail –...Always, always, write a brief conclusion of your analysis, even if your conclusion is “this approach is ...ideal because…” Including a conclusion for each analysis or task will keep you from repeating your work...

  30. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    Published
    Jan. 31, 2019, 1:32 p.m.
    ...of?  A tumor’s genesis is divided into 3 phases: initiation, progression, and metastasis. The microenvironment...altered fibroblasts and promote the development and metastasis of tumors through inciting inflammation. Speaking...microenvironments affect tumor spread and drug resistance? Although we’ve seen how multiple types of cells...story. It’s essential for a tumor to spread and to resist drugs. And to do that, let’s take a look at exosomes... case, exosomes may transport miRNA from drug-resistant cells to sensitive ones. Though this process is... expression of TAK 1, an enzyme that controls apoptosis and regulates transcription. Without TKA1, cells...cells proliferate and cannot undergo apoptosis, thus multiplying uncontrollably and building up tumors....

  31. Modulate the Activity of 17 Signaling Pathways with One Kit!

    Type
    Blog Post
    Published
    March 22, 2016, 2:30 p.m.
    ...involved in drug resistance Wood wanted to find out which particular pathways drive resistance to specific ...relatively short amount of time, cancers become resistant to drugs, particularly targeted therapies,” said...comprehensive understanding of how cancers become resistant, an emerging theme is that they do so by activating...proliferation, survival, differentiation, and apoptosis. For each of these 17 pathways, they identified... a time, you could identify those that confer resistance to drugs,” in a rapid, straightforward, and scalable... for use in screening for pathways that drive resistance to drugs in vitro, and they’ve shown that it ...confer any phenotype of interest, not just drug resistance,” he said. “It could [be used to screen for] ...

  32. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...Updated mini-transposon vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab...mini-Tn5 vector that can be used to generate random mutagenesis libraries or to insert heterologous genes, reporters...this vector, called pBAM1, by conducting random mutagenesis in the soil bacterium Pseudomonas putida and ...constitutively expressed, reports on the balance of biogenesis and mitochondrial degradation in vitro and in...the complexity of assembling recombination of transgenesis constructs? A new cloning system combining the...maximum of 24 elements assembled into a final transgenesis construct. Plasmids constructed using Golden...recombination template vectors, to perform multiple site mutagenesis and create complex fusion or recombination vectors...

  33. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...from Addgene consists of two plasmids: pCas9: carries Cas9 and chloramphenicol resistance pCRISPR: carries... targeting the gene of interest and kanamycin resistance E. coli carrying the phage recombineering machinery...recombineering is unsuccessful and the wild-type sequence persists, Cas9 will cleave the gene of interest, inducing...Insertion of Transposable Elements by Guide RNA-Assisted Targeting) and the Zhang lab's system called CAST... enable gRNA-directed transposition. INTEGRATE consists of four major components including (1) a CRISPR...Streptococcus pneumoniae. Robert Husson lab's M. tuberculosis CRISPRi plasmids A group from the Husson lab... used CRISPRi to study essential genes in M. tuberculosis. dCas9 and a gRNA are expressed on pRH2502 and...

  34. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...of plasmid multimers. Capillary Gel Electrophoresis (CGE) analysis Another technique that can detect multimers...Levene SD. Analysis of DNA topoisomers, knots, and catenanes by agarose gel electrophoresis. Methods Mol...stability study and topology analysis of plasmid DNA by capillary gel electrophoresis. BioProcess Int. 2006 ...the supercoiled plasmid monomer, and at <6 kb, consistent with the size of a supercoiled plasmid dimer.... multimers is capillary gel electrophoresis (CGE), which has been used in plasmid vaccine and gene therapy...Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med. 2020;20(10):798-805. doi: 10.2174... He Y, Lacher NA, Ruesch MN. Capillary gel electrophoresis with laser-induced fluorescence of plasmid ...

  35. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ... Blog Plasmids 101: Antibiotic Resistance Genes Antibiotic Resistance: An Old Solution but a New Problem...environments. The genes encoding these functions form the basis of many molecular biology tools! Figure 1...natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense Genes allowing...its host. For example, cells with a penicillin resistance plasmid can grow in spite of a hostile Penicillium...fungus trying to kill them off. While antibiotic resistance-mediated selection is indispensable for cloning...cloning work, antibiotics and plasmid-encoded resistance to them are the tools of a millennia-old microbial...like colicin typically also carry the necessary resistance genes to help cells survive their own secretions...

  36. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    ..., making them ideal for studying vertebrate embryogenesis. To illustrate some of the reasons that zebrafish...in developing embryos are available. Additional analysis tools have been developed to help parse and visualize...1990’s, two revolutionary large-scale random mutagenesis screens produced ~1,500 zebrafish mutants that...to study development and disease. Additional mutagenesis projects have since been initiated and today ...gene deactivation tools for zebrafish Random mutagenesis screens provided a major toehold into understanding...the zebrafish is an ideal system to observe embryogenesis live. The origins, divisions, and movements ...tracing defined a high-resolution map of early embryogenesis in time and space, and was also used to precisely...

  37. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    Published
    April 2, 2019, 1:44 p.m.
    ...many contexts within the fly including oogenesis and cytokinesis. Why name this protein after a vegetable...hedgehog exhibit abnormal segmentation during embryogenesis. Typically, regularly spaced denticles (bristles...such as cell movement/motility, development, cytokinesis, and muscle contractions (think skeletal muscle...monster, a meiotic arrest gene required for spermatogenesis One of our smaller conference rooms is named... meiotic arrest gene that is necessary for spermatogenesis. Flies that lack cookie monster are sterile...to a failure of primary spermatocytes to enter meiosis. Testis of cookie monster mutant flies are filled...White-Cooper, 2003). Why call a gene required for spermatogenesis after a Sesame Street character? The scientists...

  38. RUBY-Red Siliques

    Type
    Blog Post
    Published
    March 1, 2022, 2:15 p.m.
    ...antibiotic/herbicide resistance and fluorescent proteins. Selection using resistance genes usually involves... editing, transformation may be identified by resistance to an antibiotic. Successful editing of transformants...occurs in T0 plants (from callus) or T1 plants (Arabidopsis floral dip). In the next generation the CRISPR...planning to use it to simplify my work with an Arabidopsis mutant that can’t set seeds. I study the development...and a spontaneous step. (B) RUBY is a reporter consisting of these three enzymes expressed under the same...   Fig. 3: (Top right) A RUBY-expressing Arabidopsis seed following floral dip transformation next...without antibiotics. (Top left) Mature silique (Arabidopsis fruit) with fertilized developing seeds from ...

  39. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ...of a homologous donor (e.g., sister chromatid, plasmid, etc.), and consists of three pathways: classical...initiates primary synthesis from a template followed by a round of lagging strand synthesis to fill in the...for strand invasion and a primer for DNA repair synthesis. The ssDNA strand then displaces one strand of...classical double-strand break repair (DSBR), synthesis-dependent strand-annealing (SDSA), and break-induced ... template to serve as a primer for DNA repair synthesis, ultimately leading to the formation of double...manner, this results in a non-crossover event.  Synthesis-dependent strand-annealing (SDSA) pathway SDSA... chromosomal aberrations. Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination...

  40. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...eliminating stringent factor Allows RNA synthesis in absence of protein synthesis Ptrc-ccdA   Propagation of ccdB-containing...of 30S ribosome Confers resistance to streptomycin Tn10   Confers resistance to tetracycline Additionally...uptake, iron uptake receptor mutation. T1/T5 Phage resistance gal Mutation in galactose metabolism pathway...only gyrA, gyrA96 DNA gyrase mutation Confers resistance to nalidixic acid  hsdRMS hsdR(rk-, mk+) Unmethylated...constitutive expression of genes for deoxyribose synthesis Allows uptake of large plasmids hee   High electroporation...source for growth proAB mutation in proline biosynthesis pathway Requires exogenous proline source for...Table 2: Lab strains of E. coli Strain Natural resistance Primary Use Genotype ccdB Survival 2 T1R   ...
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