We narrowed to 747 results for: cat.1

... to the use of multiple antibodies. Figure 1: Overview of using Sandwich ELISA to quantitatively...the most common type of competitive ELISA is a modification of the sandwich ELISA as described here, the...ELISA, the only difference being the additional amplification step, which helps to visualize low-abundance...

...conclusions.  Figure 1. Potential points where DNA errors can be introduced, indicated by exclamation marks... utilize an array of tools that rely on DNA amplification and manipulation to produce and edit plasmids...places where DNA is being created, such as PCR amplification or bacterial growth. Created with BioRender....through the QC pipeline to resolve any issues.  Catching and addressing QC issues Addgene’s QC team has...has developed a process for identifying, categorizing, and resolving a variety of QC issues. This process...discrepancies can be caught prior to distribution. Catching these errors prevents future experimental issues...during analysis. These include topics such as: locations in a plasmid that are less likely to affect function...

...supervisor, even if he or she is a difficult person: 1) Understand your supervisor’s expectations Every ... series.  “The only healthy communication style is assertive communication.” - Jim Rohn, Author, Entrepreneur...I heard about supervisors in all of the above categories. In addition, I also learned about two other ...Assertiveness? An assertive person is able to communicate their ideas confidently while also considering...of the most frequent sources of conflict is miscommunication. For example, you might misunderstand your...the wrong direction. The best way to avoid miscommunication is to follow up after every meeting with an...workplace, these strategies will help you to communicate with confidence earn respect from your supervisor...

...basis of many molecular biology tools! Figure 1: Common functions of natural plasmids. Created with...Shareck et al., 2004). Plasmid categorization is complex Plasmid classification has shifted over the years...us! A plasmid is a small piece of DNA that can replicate independently from its host’s chromosome. Although...genes between bacteria can also have serious implications for public health. Bacteriocin plasmids: microbial...poisoning, the difference is virulence plasmids! This category includes genes that help bacteria invade hosts...years, and new classifications are still being created as whole-genome sequencing becomes more relevant (... common plasmid features, such as origins of replication and promoters, check out our other Plasmids 101...

...treatment and improving patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma transposon...Mechanisms of epigenetic modifications The main mechanism of epigenetic modification is chromatin remodeling...literally means ‘on top of the genome’). Epigenetic modifications can result in phenotypic changes without any...any change in genotype. Epigenetic modifications play a large role in turning genes ‘off’ and ‘on’, signaling...two main mechanisms, either through histone modifications, which can influence the transcription of genes...hypomethylation, on the other hand, has also been implicated in the development and progression of cancer ...enzyme digestion, DNA adenine methyltransferase identification, and bisulfite sequencing. However none of ...

...If you already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise ...well as suitable applications. Expression System Components of System Application Mammalian expression...CRISPR mammalian expression systems, the typical applications for each, and potential delivery methods.   ...Select the appropriate CRISPR tools for your application: Wildtype Cas9 or Cas9 nickase are appropriate... or introducing mutations and tags, while a catalytically dead dCas9 can be used in conjunction with activator...or more sgRNAs and Cas9 are required for any application. If you want to include a homology directed repair...Cas9-gRNA complex and delivered to cells using cationic lipids. Transient expression of CRISPR components...

... – a mechanical device that could be used to bend 1/8th inch steel wire into a backbone structure of a...development programs for bioscience educators. Using these hands-on tools, educators introduce their students to...biologists have always struggled to effectively communicate the nuances of protein structure in a flat 2-...dangerous. Subscribe to CRISPR Blog Posts How do educators and students use molecular models?   We created...something tangible. Under the guidance of a trained educator, these instances of focused attention become magical... their labs. Soon after we began working with educators in our Center for BioMolecular Modeling, we spun...through flexibility It is common knowledge in the education community that All models are wrong….Some models...

...viruses - 2 cas systems regulate at the DNA level and 1 at the RNA level. Udi Qimron from Tel Aviv University...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...Guangzhou and the Science City The conference was located in the modern, relatively new “Science City” area...and technology companies and start-ups choose to locate there. I made time to go into the city of Guangzhou...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...Steitz quipped that RNA in the ribosome is in a complicated conformation “not like DNA which has a boring...

...which you’ve already fluorescently labeled. Figure 1: Using Split-BioID to Identify Protein Protein Interactions...methods for identifying PPIs, such as affinity purification mass spectrometry (AP-MS), lack the ability ...the validation of a binary PPI as well as the identification of additional interacting factors. Problems...detect weak and/or transient PPIs. Many PPI identification techniques rely on protein interactors to stay... AP-MS. This requirement often prevents the identification of weak and/or transient interacting proteins...spatiotemporally defined protein complexes. Nature communications. PubMed PMID: 28585547. PubMed Central PMCID...

...is best for larger genes such as Cas9. See figure 1 for an example of how changing transfection ratios...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from...end-users, size of the transgene, and downstream applications should all be considered when choosing the correct... as resources available, cost and downstream applications. For example, while the PEG precipitation method...

...significant progress in clinical trials.     Figure 1: Types of delivery and editing strategies for CRISPR... therapeutic potential in mice with a dominant cataract disorder caused by a single copy of a gene known...known as Crygc (Wu et al). They found that the cataracts phenotype could be rescued by zygote co-injection...substantial relief from both disorders. As of publication, submissions to the UK, EU, and USA federal drug...10.1186/s12943-022-01559-z Zarghamian P, Klermund J, Cathomen T. Clinical genome editing to treat sickle cell...

... inhibitory synapses in the brain.  Figure 1:Schematic diagram of different FingR constructs and... labeling proteins with different subcellular locations. Image from Goudeau et al., 2020. Goudeau... show a high proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in ...in high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...Hybrid for Multiplexed Editing and screening Applications (CHyMErA) Knockout Libraries express hybrid ...

...). References 1. Li, Chaokun, et al. "FastCloning: a highly simplified, purification-free, sequence-...methodologies, with a special emphasis on these five categories. Please note that many of the following cloning...relies on site-specific recombination reactions catalyzed by BP Clonase and LR Clonase enzyme mixes. After...there is no need for restriction digests, gel purifications, or ligations. The general Gateway cloning protocol...

...article, they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...BioRender.com In their recent ACS Synthetic Biology publication, Mary Dunlop’s lab generated a photosensitive...regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd dimerizes bringing the C-terminal...

...SpCas9 base editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated...Lower on-target activity than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3  HF-Cas9 (NGG)  Very ... plasmid or group of plasmids' functions and applications. We hope that these articles make it easier ...FLIM (Fluorescence Lifetime Imaging Microscopy) applications. To demonstrate the power of mCyFP1, the Lin... Coselection efficiency is so high that some applications may permit working with edited “pools” of cells...showed high editing efficiencies with AsCpf1, indicating that ouabain coselection is a robust and flexible...be found at Addgene and in Figure S12 of the publication. Agudelo D, et al. Nature Methods. 2017. PubMed...

...technologies (Bradbury & Plückthun, 2015a).   Figure 1: (Top) Polyclonal antibodies are produced in animals...cells, for antibody production and subsequent purification.  Academic laboratories and companies have already...Edition. 2104. Cold Spring Harbor Laboratory Publications, New York. Helma J, Cardoso MC, Muyldermans ...

...most comprehensive for searching. Timesaving Tip #1: If you know the species that your sequencing result...search to check for expected ORFs, mutations or truncations. A primary advantage of blastx is that you do...other labs speed up their plasmid and cloning verification steps and free up more time for using your plasmids...

...genome engineering and microbiology.     References 1. Chen, Sean, et al. "Highly efficient mouse genome...valuable to scientists, there are some potential complications that must be considered when using this method...alternative approach, which avoids many of these complications, is to directly deliver a ribonucleoprotein ...used for most of the current genome engineering applications of CRISPR including: generating single or multi-gene...pathways. This makes Cas9 RNPs useful for CRISPR applications where limited expression of Cas9 is required...development. David Liu’s Lab has demonstrated the use of cationic lipid-mediated methods to deliver Cas9-gRNA RNPs... of eventually using this method for in vivo applications. This technique utilizes Cas9 proteins harboring...

... questions of how the brain operates. References 1. Callaway EM. Transneuronal Circuit Tracing with Neurotropic...able to replicate, and for a virus to cross a synapse from one cell to another, it must replicate. Modified...like how our stem cells are housed in specific locations and give rise to progenitors during growth and...infects one cell (the “starter cell”). It then replicates within that cell, and nascent RABV cross synapses...infection occurs, subsequent cycles of viral replication and retrograde transmission occur, spreading ...limitations. First, these high levels of viral replication are toxic. Second, infecting subsequently higher...spread monosynaptically from these cells. These modifications take advantage of the fact that RABV is an enveloped...

...in restraint. The very first subheading in Chapter 1 asks “What is science?” That’s a very big question...drives and their ramifications. Readers unfamiliar with academia’s challenges with publication and funding ...Connecticut College who has been teaching and communicating science for over two decades, attempts to answer... primarily on fluorescent proteins and their applications, tackles much broader issues in this brief volume...sexism in science, problems with peer-reviewed publications and funding, bioethics in the age of CRISPR,... and her essential-but-unlauded role in the identification of DNA’s double helix. Perhaps to the dismay...dismay of middle-aged scientists, the identification of Green Fluorescent Protein (GFP) and its development...