We narrowed to 988 results for: des
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TypeBlog Post...individual molecules. Poor photon budgets are a widespread issue with fluorescent proteins—even the thriftiest...student searching couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical... the complexity of the system. A particularly desirable property for small-molecule labeling strategies...size, brightness, and cell permeability. In past decades, further refinements to this established dye structure...spectral range. Nevertheless, these fluorophores were designed as antibody and oligonucleotide labels for fixed-cell...
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CRISPR Antimicrobials
TypeBlog Post...beta-lactam resistant population to the antibiotic, despite this resistance gene being encoded by a high-copy...currently undergoing a renaissance. Yosef et al. designed a two-phage CRISPR system to resensitize antibiotic-resistant...immune system, may someday be used to target the hardest-to-kill bacteria. In using phage-derived vectors...Ahmed A. Gomaa, and Rodolphe Barrangou. “A CRISPR design for next-generation antimicrobials.” Genome Biol...Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design Tools ... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog Post...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...cytosine and adenine base editing, called CABE-Ts and TadDEs without additional fused components (Lam et al....editors have a narrow editing window of only a few nucleotides, so they are tightly constrained by PAM availability... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog Post...manner. These videos can be especially useful when designing new primers to clone your sequence of interest... your cloning. Most sequence analysis software includes Golden Gate cloning tools that simulate the restriction–ligation...according to their concentration, size, and the desired insert-to-vector ratio using the equation in Figure..., positive colonies may contain plasmids with undesired ligations, so always screen multiple colonies ...see in colony PCR electrophoresis), is free of undesired mutations, and that you have the correct four-... -
Hot Plasmids Spring 2024
TypeBlog Post...1: CROPseq-multi uses two sgRNAs with internal barcodes (iBARs), multiplexed using tRNAs, within the lentiviral...separation of the multiplexed elements. The team designed sgRNAs that include an internal barcode and separated... have been made since prime editing was first described in Anzalone et al., 2019, including modifications... sparsely-labeled field of view that was more desirable than having many closely-packed labeled cells.... combination with miR124T targeting sequences, provides a simple yet valuable tool for microglia research... -
Antibiotic Resistance: An Old Solution but a New Problem
TypeBlog Post... defined as microbial survival despite exposure to antibiotics designed to kill them or to impede their...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...different reasons. Exposure to antibacterial drugs provides a selection pressure, killing off bacteria that...equipment, etc. Virtually any preventive measure designed to help keep you from getting sick can be helpful... -
5 Reasons to Use Reddit for Science Communication
TypeBlog Post...They also host AMA forums where users can ask designated scientists anything about a particular topic....calendar invite over to the r/Science team with a description of our AMA and Joanne’s preferred time to start...was writing all day and straight on through her designated two hours for answering questions. She was able...toxic advisors, fruitless job searches, and even despair at having chosen an academic past. I hope I was...receive via email. The cool features are too many to describe here, but I have over 2,000 notes which Evernote... -
Analyzing CRISPR Editing Results with ICE from Synthego
TypeBlog Post...Engineering Solutions, which also includes our CRISPR knockout guide design tool, guide validation tool, and...their relative abundances. The ICE tool also provides a Knockout score (KO-score) which represents the...the CRISPR workflow. For a multiplex sample, ICE includes visual representations of all detected edit types...Contributions, Indel Distribution and Traces - provides particular details about the indel profile of ... -
Deciphering the Mysteries of Behavior with Viral Vectors
TypeBlog Post...Over the past decades, neuroscientists have made great strides to map brain regions and allocate specific...for this task. However, most lesioning methods destroy areas larger than intended and may unintentionally...surface. The latter reflects a state of behavioral despair, and the more time the animal spends in the passive...that were not (Hare et al., 2019). The research described here is only a small selection of the increasing... -
CRISPR-mediated Plant Base Editors
TypeBlog Post...second outcome and increases the generation of the desired T-A base pair. Figure 1: Transition mutation...by base editors. Pathways that compete with the desired outcome are not pictured. Adenine base editors... SpCas9-VRER TadA-TadA* NGCG 8 Tips for designing base editing experiments in plants Choose a base...successful base editing causes the creation or destruction of any restriction enzyme (RE) recognition site... indel sometimes may also results in RE site destruction. Many thanks to our guest blogger Kutubuddin... -
Tips for Using FRET in Your Experiments
TypeBlog Post...data from your FRET experiments. You may have to design several different constructs to find one that could...studies. These parameters are less important when designing a biosensor, as the distance between the two fluorophores...gene of interest: Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine... straightforward and quantitative, but it is destructive and cannot be used for dynamic measurements. .... Extra care should be taken so as not to destroy the donor molecule. Fluorescence lifetime imaging microscopy... -
Plasmids 101: The Promoter Region – Let's Go!
TypeBlog Post...insert into RNA. In practice, the term "promoter" describes the combination of the promoter (RNA polymerase... Primarily used for RNA transcript Description Expression Additional considerations CMV General... phosphoglycerate kinase gene. Constitutive Widespread expression, but may vary by cell type. Tends ...promoters Promoter Primarily used for Description Expression Additional considerations T7 in ...identify the right tissue-specific promoters as described by researchers at the NIH here. Note: A. Max... -
Antibodies 101: Beyond Surface Labeling
TypeBlog Post...when it comes to fluorophore selection for panel design. Find streptavidin plasmids and biotin plasmids... this system is increased flexibility for panel design. Another perk is that one primary antibody can ...for example, a previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside analogs...walk through the topics we covered in this post. Besides direct labeling, indirect labeling can be used ...surface, dump gating is a useful way to get rid of undesired cell populations. Dump gating uses the same color... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog Post...recombinant antibody. They are ~25 kDa single polypeptides that contain the variable light chain (VL) and...lethal to animals. In the clinic, scFvs’ size also provides advantages over antibodies (Ahmed et al., 2012... bond between the VL and VH domains. This bond provides stability and solubility for the scFv (Gaciarz...clinic (Bates and Power., 2019).Many of the scFvs designed for oncology applications are reformatted as bispecific...Chantima W, Sangma C, Poomputsa K, Dharakul T (2015) Design and Generation of Humanized Single-chain Fv Derived... -
Antibodies 101: The Basics of Western Blotting
TypeBlog Post...protein analyses using ELISA. Peptides 72:4–15 . https://doi.org/10.1016/j.peptides.2015.04.012 Burnette WN ... Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode/iblot ...blocking buffers, and incubation times. If the vendor includes publications that cited the antibody, consider...abundance In addition, the thickness of the band provides information about the relative abundance of the... -
Technique: Probe Phage Genomes for Host Binding Proteins
TypeBlog Post...identify possible gene function. In this post, I will describe a useful and cost-effective method that our lab...and probe with live cells The following is a description of a technique published by Simpson, Sacher, ...ends) and ligate into a bluntly digested vector designed for protein overexpression in E. coli, such as...the original plate to identify which phage gene encodes that protein! First, block your membranes with ... can lead to high background that obscures the desired signal. We have successfully corrected this by ... -
Transferable Skills Guide: Teamwork
TypeBlog Post... the work, shared the credit, and we both won accolades that would launch our careers in science. Fast...helped assemble a crack team of Addgenies that includes: Michelle, the Senior Scientist who coordinates... a happier person. What does a good team need? Besides that all-important sense of purpose, there are ...Slack constantly. Whether your teammates are two desks away or on the other side of the world, communication...self-interest ahead of the team’s goals, then that team is destined to do poorly. To some extent, it doesn’t even ... -
Addgene Stands with the Scientific Community
TypeBlog Post...critical research infrastructure that has taken decades to establish.These funding decisions will have ...community for years to come. We Are Here to Help Despite these challenges, Addgene remains steadfast in ...educational resources: Our eBooks, videos, science guides, and protocols remain freely accessible to all ..., helping teams to be successful in their work despite limited resources. These resources become even ... -
Troubleshooting and Optimizing a Western Blot
TypeBlog Post...right. If you’ve thoughtfully done the technical design of your blot, but are still finding yourself having...want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to make sure you have the...your membrane! Membranes look the same from both sides, so it’s quite easy to think lane 1 is lane 8. ... with smaller tasks, or simply enjoy some well-deserved down time while waiting for your timer to ding...things you’ll want to consider (see our “technical design of a western blot”) post, most imaging issues can....2011.06.001 More resources on the Addgene blog Technical Design of a Western Blot More resources on addgene.org... -
Your Top Requested Plasmid in 2016!
TypeBlog Post... and the Puromycin resistance gene as separate peptides. PX459 This is the vector backbone. This particular...Protocols Paper for more information). V2.0 This designation separates pSpCas9(BB)-2A-Puro (PX459) V2.0 from... the Zhang Lab's Nature Protocols Publication describing them here. References 1.Ran, F. Ann, et al. ...Guide Pages Browse Our CRISPR Collection Find gRNA Design Tools ...