...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...

... 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...

...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...

...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...

...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...

...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...

...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...

...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...

...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...

...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 ...

...Open Bioeconomy Lab (Link opens in a new window) designed and codon-optimized the Open Enzymes parts for... Open Bioeconomy Lab's crowd-sourced expression guides (Link opens in a new window) . Open Enzymes are...

... 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 ...

...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 ...

...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...

... 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 ...

...select the optimal dye or fluorescent ligand when designing an experiment. You may find yourself defaulting...protein, enabling the labeling of protein substrates/peptides and antibodies. Example applications of this include..., which is an important consideration for assay design (Breen et al., 2016). When selecting dyes, it's...fluorescent donors and acceptors in their product descriptions, or technical manuals. Specific applications...signal-to-noise ratios. Certain dyes have been specifically designed for use in super-resolution microscopy techniques...

...below. Plasmids ID Plasmid Description PI Viral Preps ID Viral Prep Description PI (Cargo) PI (Capsid) 214869...

... producing a wall of results that can only be described as bigly. But such a project should not be undertaken...Library Screening Why the kinome? While properly designed perturbations – in the case of screens using CRISPR...References: 1. Doench, John G., et al. "Optimized sgRNA design to maximize activity and minimize off-target effects...with CRISPR/Cas9 Get John Doench's Tips on gRNA Design Check Out News Ways to Use CRISPR for Single Base...

...lab earlier, to read that stack of papers on your desk, and to tackle some new experiments before the next...gathering that pile of lab notebooks and binder on your desk. Arrange them in chronological order (or by project...labels. Color code your Post-its and write brief descriptions on each. Place the notes sticking up out of ...colleagues. Start small by identifying the protocols, guides, or references that your labmates ask to borrow...

... the parts of CRISPR. The bacterial chromosome encodes a tracrRNA (in some systems including Cas9), Cas...DNA sequences. In laboratory settings, an sgRNA includes the crRNA and tracrRNA sequences in a “single-... sequences in the CRISPR array that will become guides for future targeted degradation. Before integration...biology of Cas9. Want a simpler guide to keep at your desk? We’ve got a CRISPR cheat sheet for that! Resources...