We narrowed to 373 results for: proc.2

...go anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes... the technology. This transition takes around 1 – 2 years. The product is the measure of success. Technology...time to speak with us about the biotech startup process and work environment! Interested in Synthetic Biology...

...animal is given additional booster immunizations every 2-3 weeks to increase the antibody titer. The titer ...level of antibodies is reached - typically within 2-4 months. After the animal has the desired level of... - which contains the antibodies - is further processed to isolate the antibodies. Most commonly, antibodies...

...short-read sequencing are highlighted in Figure 2. Figure 2: Overview of short-read sequencing technology...overview of the QC process at Addgene. All plasmids go through this same initial QC process using an Illumina...script can then continue in the auto-align process. This process is typically reserved for deposits that ...the highest quality and accuracy during the QC process and therefore put in a lot of effort to stay up-to-date...sequencing technologies that Addgene uses in the QC process, when these technologies are used, and how Addgene...first widely used DNA sequencing technology. The process relies on “sequencing-by-synthesis,” where each...by synthesis, but modifies the original Sanger procedure to improve throughput by increasing the number...

...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...arise very quickly following the tried and true process of evolution. Watch this video showing bacteria...targets an essential, conserved, non-negotiable process within the bacterial cell, antibiotic resistance... the development of resistance; it’s a natural process of life and one which has allowed bacteria to evolve...

...and the Journal of Emerging Investigators. Chapter 2 - Getting high school students and teachers interested...networking!) and the manuscript review/submission process at JEI. Chapter 3 - JEI target audience and their...

...this quick trick. Just flip, and press! Lab tip #2: Tube flick trick When combining reagents in the lab... time, our lab has gotten a lot experience in processes including automated vial filling, accurate tube-labeling...

... anniversary! We've done so much in the past two (2!) decades that it's hard to summarize, but we wanted...support to our efficient and easy tech transfer process to our website and educational resources. We've...

...form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines at promoters and...pX330a dCas9-LSD1 for targeted removal of H3K4me1/2 and H3K9me2 histone modifications. Like the TALE-LSD1...approach for targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids... and targeting the epigenome may make certain processes, like cellular reprogramming of iPSCs, more efficient...

...certain states of interest. For example, in Figure 2 below we present a two-input, five-state GRSM register...linking RSM inputs to endogenous gene expression (2). Our paper (1) explains how to arrange attB-attP...Science. 353, aad8559 (2016). PubMed PMID: 27463678. 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...used to better understand and engineer cellular processes that are influenced by temporally ordered biochemical...

...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...might feel disconnected from the entire scientific process.  I've spent a lot of time debating how to fix ... science in May, I marched to advocate for the process we use to learn about the world. In a way, I marched...

...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...selection process to standardize our feedback and evaluation of viral vectors. This process builds on ...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific experimental...its inherent bias: Addgene uses a prioritization process to decide which tools we make available. This ...us first identifying broadly useful vectors, a process that isn’t as simple as it seems. Take, for instance... and blog posts. The lessons learned from this process have shaped and supported our growth and development...

...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...control point of DNA replication in bacteria, so the process is carefully regulated by a mix of positive and...replication (oriC), where initiation begins. This process relies on the initiation protein DnaA. First, DnaA... versions is required to start the replication process. When enough copies of ATP-DnaA are bound, including...integration host factor (IHF) assists with the process: it bends the DNA to redistribute the DnaA from...plasmid ori, after which replication of the plasmid proceeds (and terminates) similarly to the bacterial chromosome...

...residues, which can be found throughout the antibody. 2) Endogenous cysteines, such as those that make up ...Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 ...yourself, there are kits available to make the process quick and easy. Before getting started, be sure...

...studies have shown that the conditions Sotos-type 2 and Marshall Smith Syndrome are due to mutations in...accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith) and could be expanded to NFIA-...lines and use these lines to study how immune processes and biological pathways are altered in PMM2-CDG...

... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...high background or low signal during the imaging process, try adjusting your exposure time first, since ...

...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I .... During the propagation and/or transformation process, some bacteria will either never receive a copy...antibiotics listed below affect continuous cellular processes (e.g., protein synthesis), while amp only affects...

...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression..., studying the genetic underpinnings of these processes has been complicated by the lack of precise genetic...undifferentiated cells at the end of the screening process. However, while the neuronal marker TUJ1 is useful...

...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...to immunoprecipitation and simply refers to the process of anchoring an antibody, often called the capture...protein interactions, and limit high throughput procedures. However, agarose beads are often a more cost-effective...

... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...have been cross-adsorbed. The cross-adsorption process filters out off-target antibodies increasing specificity...

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...decrease false positives observed in the cloning process, making it easier to produce the constructs you...novel and powerful pyranopyridine derivatives." Proceedings of the National Academy of Sciences 113.13 (2016...-time observation of metabolite production." Proceedings of the National Academy of Sciences 113.9 (2016...pCri system to simplify the protein purification process by making it easy for a researcher to quickly generate...on the system Easing the Protein Purification Process with pCri Cell-free protein expression kit Cell-free...

...break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases indel formation... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...the template and primer binding site, increase processivity, or improve thermostability. What came out on...repair (MMR) complex, to temporarily inhibit this process and promote copying of the edited strand to the...to the numerous innovations that streamline the process of prime editing, the Liu lab contributed an improved...

...whether the results include knockout data.    Figure 2: Search options in the Antibody Data Hub. Data reports...antibody worked in your application. The submission process is easy, with simple, descriptive fields, drop-..., and can be saved at any point throughout the process, so you can work through it at your own pace. To...

...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...activation across multiple loci was an arduous process. When using zinc finger proteins or TALE proteins...

... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...PMID: 24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2...volume. Always remove packaging cell debris before proceeding to ultracentrifugation to avoid contaminating...

...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate... reduce recombination during the amplification process. In liquid culture, all of the bacteria can grow...

... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...MMEJ doesn’t get a lot of press. However, this process accounts for a percentage of the mutations seen...

...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives... enabling the study of dynamic, physiological processes. Re-engineering GFP to increase its range of color...

...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...others for failing to find our work?  I study processes for diagnosing and resolving organizational performance...impact of their contribution. It offers a 5-step process and several examples for creating effective research...

...citizens enter into in order to protect their rights (2). Less formally, governments are the organizations...science education and literacy that this changes.  2) Lumping all scientists (and consequently their results...politics? Science is an apolitical process for seeking knowledge. The process begins with a testable hypothesis...hypotheses and the cycle begins again. No part of this process is inherently political so why is there a march...scientists as well as other citizens. Thus civilization proceeds in an informed fashion. For science to be useful...

...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...looking for, you are ready to begin the screening process. Want to set yourself up for a screening success...then a single, bulk validation may be enough to proceed with your experimental plan. Validating knockout...

...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...suggestions for this year’s teams that may be procrastinating choosing their costume?  As our organization...

...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...of optogenetic control of endogenous cellular processes." The LITE system can now be applied to unraveling...genetic and epigenetic players in normal biological processes and disease states. While LITE plasmids have clear...

...would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for...gating on the negative population.     Figure 2: Dump gating labels all unwanted cell lineages with... for entry of the antibodies. Here, a two-step process is employed: the cells are first fixed and then...phosphatases, and ubiquitin ligases; and many more. This process, however, leaves the nucleus intact. If markers...not need to use a separate fix/perm cytosolic procedure in that case.     Figure 3: For intracellular...

...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic...great way to stimulate and study these physical processes. Joachim Goedhart and colleagues in the Gadella...could be applied to study other actin-dependent processes in cells and tissues. And, this Opto-RhoGEF design...the protein of interest supports a variety of processes in different cell types. One great way to do this...

...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...allowing them to optimize the compound production process. How have you used FP timers? Are you planning ...

...your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link biomolecules...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ....)  Step 5: Image   Figure 1: The ExM process allows you to physically magnify a sample so you...

...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...tracking software. Locomotion is a highly complex process on the level of individual neurons and in terms...

... a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using...expensive system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of...gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences...  Figure 1: Overview of the Western blot process. The sample, a mixture of protein is separated ...antibodies, and finally visualized.   This process allows you to detect a single, specific protein...conduct the electric current. The wet transfer process is very effective in transferring a broad range...common readout for subcellular fractionations, a procedure that isolates proteins from different cellular...

...bringing in environmental contaminants.  Figure 2: An Addgenie works in the tissue culture hood while...doi.org/10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma ... enter the culture medium during subculturing processes or from the air if the cell line remains exposed... infecting mycoplasma. The primers used in the process are either species/gene-specific or universal. .... Some amount of expertise is required in this process because result interpretation might be difficult...by training eligible staff for the cell culture process and hence eliminate one of the causes of contamination...filtration. There is no 100% assurance that this process can avoid contamination but it’s a very good practice...

...Chapter 1 - Who are Addgene's Founders? Chapter 2 - The process of founding Addgene, where the idea for Addgene...

...Almo, Institute for Protein Innovation  1:18:45 - 2:01:18 (end) - Panel Alex Tucker (Ginkgo Bioworks)...their curation of citable, linkable figures. Cause #2: Hidden “negative” data We all want to be able to ...doing an experiment wrong. However, even simple procedures like pouring a plate or running a gel have intricacies... retain searchable, thorough notes on how lab procedures were actually performed, potentially curtailing... sense on many different levels - it’s hard to process large amounts of information and, from a SciComm...how and why papers change throughout the review process. This will hopefully push reviewers towards more...experimental design as part of the publication process (Nosek et al 2018). In this ideal situation, a ...

...phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2 coat... 296.5569 (2002): 913-916. PubMed PMID: 11988576. 2. Tsien, Roger Y. "The green fluorescent protein." ...for biological spectroscopy and microscopy." Proceedings of the National Academy of Sciences 114.23 (2017... real-time multicolor luminescence imaging." Proceedings of the National Academy of Sciences (2015): 201418468...

...tagging." Nature methods (2015). PubMed PMID:26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and...the attributes of the native protein. Since this process takes a lot of time and energy, Martí Aldea and...

... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ...which we've discussed in two follow-up posts: (1) (2). Base editors have a narrow editing window of only...repair. Black arrows indicate natural cellular processes. BER = base excision repair. Note: hypoxanthine...

...Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One Step, Precision...contributing more modules into the public domain in the process. “We had several motivations for making GreenGate...

...having trouble preparing your most excellent resume. 2) Follow job boards and check them regularly Use your...existing relationships as part of the job application process can dramatically increase the chances of being ...document. 6) Use your network! During the entire process, use whatever means you can to get your resume ...

... 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse experiment...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA...not always guaranteed. To learn more about this process check out our Mouse Modeling blog posts: Part 1...have germline competent ESC lines. Second, the process is much quicker. It takes 3 months to generate ...

...photoconvertible fluorescent protein. Nat Methods. 2009 Feb 6(2):131-3.  PubMed. Find plasmids from this publication... be expensive and time-consuming to repeat the process with these new photoconvertible proteins. With ...state after excitation at 561 nm. This imaging process worked well with the common GFP variants EGFP, ...

...Pringle’s former lab at UNC Chapel Hill. Bähler et al.(2) describe a modular collection of plasmids for a wide...Homologous recombination is the process by which nearly all domains of life repair genomic damage, specifically...Researchers have long taken advantage of this natural process to integrate protein tags into the genomes of S...

... ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...small-molecule fluorescent labels. Nature Methods, 19(2), 149–158. https://doi.org/10.1038/s41592-021-01338...Allikalt et al., 2020). Impact of labeling process: The process of dye labeling, such as conjugation with...beneficial when studying dynamic processes like mitochondrial dynamics. This process, involving mitochondrial ...specimens has offered illumination on biochemical processes that are crucial in the world of biological research...result in cellular toxicity due to the permeation process. Advanced technologies in fluorescent dyes/ligands...