...No No Yes Yes Lab Made Competent Cells Required Yes No No No Comparison of REPLACR-mutagenesis with...mutations, large deletions/insertions require complicated, often costly in vitro assembly methods. A new...-mutagenesis also requires making your own electrocompetent cells in which the phage recombination machinery...

...degradation In the Bradner and Gray labs, we synthesize compounds that enable selective removal of proteins-of-interest...FKBP12F36V-tagged proteins. The dTAG molecule induces ternary complex formation between the FKBP12F36V-fusion and the... for degrading targets in different cellular compartments, and in vivo utility. dTAG molecules will be...lentiviral plasmids can serve as a rapid means of comparing functionality of N- and C-terminal fusions and...

...is dependent on proteasome function. TRIM-away compared favorably to drug inhibition, with TRIM-away targeting...larger cell populations, you can deliver these components together using electroporation. In these bulk...250 nm), and studying endocytosis is further complicated by the reliance on pH-based biosensors used to...rapid and reversible recruitment of chromatin complexes to any genomic locus in any cell type. By varying...

...process that’s difficult for individual labs to complete. The nanobody based RANbody platform from the ...the sample due to the smaller size of RANbodies compared to antibodies. Staining with multiple HRP-RANbodies...sample is stripped. HRP-RANbodies are, however, compatible with most multi-labeling immunohistochemical ...antigen with a primary/ labeled secondary antibody. Compatible with multi-labeling protocols when other antigens...

...show. After graduating with a master’s, Susan completed a science writing internship with the National...planning.” To beef up her resume, she decided to complete a certificate in Strategic Communications and ...the most satisfying parts of the job is having completed one of these difficult pieces: “It’s probably ...science communication] needs. I worked for one company, but there were a ton of others that also did work...

...camelid antibodies to antibody fragments to tools completely unrelated to antibodies.  Why use an alternative...entirely new affinity reagent.    Fig. 1: A comparison of an antibody (left) to several alternative ...through engineering instead of cleavage. They are comprised of the variable heavy and light chain regions,...immunogenic, but they have lower affinity and longevity compared to antibodies, along with a higher likelihood ...

... the bulk purification steps keep the cost low compared to other antibody formats. This low barrier to...is no more of that exact polyclonal antibody composition available. If you’re going to need a large quantity...antibodies produce a higher level of background noise compared to monoclonal or recombinant antibodies. This ...polyclonal antibodies for your experiment and how they compare to other antibody options out there, you’re set...

...silencing complexes (RISCs), which detect dsRNA in the cytoplasm and use it as a guide to complementary RNA ...RNA sequence. Once a compatible sequence is found (typically RNA that the RNAi is designed against), RISC...and will dissociate when gene transcription is complete. However, transcription of thousands of genes ...

... are considering to make sure that they’ll be compatible with your instruments, each other, and your target...target. I love FPbase’s Spectra Viewer tool for comparing different fluorophores. We also touched on fluorophores... sure to check that your antibody’s buffer is compatible with your conjugation kit, since many common ...reactions. If needed, you can often remove the incompatible agents by dialysis or gel filtration.    There...

...recombination events. The system consists of two components derived from the P1 bacteriophage: the Cre recombinase...recognition site. The P1 bacteriophage uses these components as part of its natural viral life-cycle, and ...and researchers have adapted the components for use in genome manipulation. Cre recombinase, originally...other sites of the same type, but are not cross-compatible. Using different lox site variants allows Cre...

... formation of replication competent lentivirus (RCL). Replication competence can arise from recombination...drive high levels of expression of certain viral components; according the manufacturer these changes result...downstream. When assessing lentiviral titer, don't compare apples to oranges After choosing a packaging system...

...optogenetic or chemogenetic receptors) enables complex control of specifically defined neuronal populations...how newly generated neurons integrate into the complex pre-existing circuit of the brain. Another study...anatomical regions of the brain receive inputs and compute outputs. In general, RABV monosynaptic tracing ...circuitry, and continue to contribute answers to the complex questions of how the brain operates. References...

... is important to fully characterize the compound, or compounds, you choose to ensure reliable results ...In order to understand the complex wiring of the brain and the underpinnings of neurological disease,...pharmacodynamic actions of clozapine-N-oxide, clozapine, and compound 21 in DREADD-based chemogenetics in mice. Scientific...

...’ll share the basic composition of these solutions below. Alternatively, companies including ThermoFisher... tool to identify a protein of interest in a complicated solution like cell lysate. But they can be a ...lost each time you strip a blot, you should not compare the strength of bands from different rounds of ...

...audience’s eye and draw people in. That can be accomplished with an exciting title and attractive figures...in less than a couple of minutes. How does one accomplish all of that? What follows are some tips to help...logical progression of your work, and you can accomplish this by employing a neat and organized format...want to make sure that the color of your font is compatible. Use high-contrast between these colors so that...

...up clear tasks for each member of the team to accomplish so that many elements of the project were able...involved definitely contributed to our ability to complete the project on such a short timescale. Our article...extensive infrastructure of molecular and electronic companion technologies that has transformed the assay’s ...of Zika Virus Using Programmable Biomolecular Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids...

...better at producing a compound of interest, thus allowing them to optimize the compound production process...fluorescent proteins have usurped old methods or complemented them in ways that make them much easier. A special...Although it is possible to make a timer system composed of two separate FPs (one that matures quickly ...

...protein capsid encapsulating the genome is majorly composed of three proteins that can be easily modified ...Delivering gRNA and Cas9 using adenoviral vectors As compared to other viral vectors, AdVs can provide a large...AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed cells yielded...vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array...

... capture large field-of-views (6). For more comprehensive reviews on several other LSFM configurations...multiplying couple-charged device (EMCCD) or complementary metal-oxide semiconductor (CMOS) sensors, it...labeling in whole tissue (15).  CLARITY is also compatible with LSFM, a method termed COLM (clarity-optimized...spatiotemporal control over labeling methods as compared to transgenic animals (17).  These viral plasmids...

...For example, Drosophila lines with compound autosomes are completely infertile when mated to wild fruit...with appropriate measures. Barriers should be a component of all gene drive confinement strategies, but ...: I. Cage Studies of Chromosome Replacement by Compound Autosomes in Drosophila melanogaster. Genetics...

...this situation: You’ve spent hours scouring a company’s LinkedIn pages, given yourself the bathroom mirror...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...the candidate has what it takes to thrive at the company and add value, not just “do the job”. When they...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...

... an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused to an RNA adenosine...off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target sites from the 18,385...edits by ~45% and off-target A-to-I edies by ~94% compared to RESCUE.   Addressing the potential use of RESCUE... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...

...orders were affected by the COVID-19 pandemic, we compared the number of plasmid orders and new COVID-19 ...the likelihood in our Poisson process into two components: 𝜃x(t) and 𝜓z(t). 𝜃x(t) describes the exponential...effects fairly well. However, we observed one complication in the China data – there is a period of 1 month... lockdown in China, then the observed data is compatible with an infinity-fold reduction in activity due...

...real time. When the PCR is complete, the results are analyzed by comparing the threshold fluorescence ...amplification reaction to take place.  When the PCR is complete, a droplet reader extracts the droplets from the...Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers ...

...his program. Initially, Provost made his labs completely open-ended, allowing every student to work on...Kowalski et al., 2016; Scott, 2021). In fact, completion of Provost’s CURE increased students’ confidence...them to near homogeneity using a 6XHis tag, and compared their specific activities by measuring NADH production...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...

...gross comparison of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids....PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived from. They also...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

...lentiviral vectors! Lentiviral vector history and components In the early 1990s, researchers developed viral...system, the possibility of creating a replication-competent virus via recombination between the transfer and...recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through the addition...Lentiviral barcoding libraries have been used for complex applications like lineage tracing in heterogeneous...

...restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less money... few ways you can do this. If you are going to complete your colony PCR analysis in the same day, you ... product size on a gel   Now that your PCR is complete, it’s time to run the products on an agarose gel... larger sized products for positive clones (+) compared to negative clones. Finally, orientation-specific...

...is ideal for tracking large collections. Many companies, such as Thermo Fisher and Micronic, provide 2D... time. 1D barcodes can be purchased from many companies such as Corning. Tubes can even have specific ...specific barcodes etched or printed onto them by companies such as Tubewriter. Any lab can benefit from using...tubes themselves. Although we’ve developed our own computer system for tracking samples, it can be done easily...

...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...give to post-docs and graduate students as they complete their appointments in your lab?  Fujii: These ...survive as a researcher because of the tough competition for academic positions and poor grant situation...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab...

...on OWW, including those for preparing your own competent cells. If you’re familiar with the process of ...service free of charge to scientists by allowing companies to pay publish their catalogs on the site. Like...of checking off each step as you painstakingly complete your experiment – one small victory in a long ...manage scientific data: Electronic lab notebooks compared with Google Docs and wikis - by Rory Macneil on...

...reaction creates an expression clone with all of the components necessary for gene expression.     The LR... are performed, the next step is to transform competent E. coli cells and select the positive clones. ...functional. Advantages of the Gateway cloning method Compatibility and flexibility Once you generate the entry ...have generated and deposited their own Gateway-compatible plasmids with Addgene. These can be used to express...

... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...in order to be expressed stably in neurons. As compared to epitopes tags, FPs are intrinsically fluorescent...fluorescent proteins (smFPs). smFPs are literally composed of FPs fused to multiple epitope tags. Indeed,... structure resembles images of the deity (see comparison image above). The church of the Flying Spaghetti...

...interior target sites. In a second step, the two compatible target sites are cleaved out, leaving behind ...a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al. 2003). Flex vectors...within neuronal networks in living animals. To accomplish such cell specific manipulations in vivo, FLEx...knockout. In this experiment it is possible to compare neuronal morphology and synaptic connectivity before...

...of DNA and both identified lymphocyte antigen 6 complex (LY6A) as a receptor for PHP.B. Read on to learn...variants that reached the CNS following IV injection. Compared to AAV9, PHP.B has a >40-fold increase in CNS ...rat CNS: AAV PHP. EB vector dose–response and comparison to AAV PHP. B." Gene therapy 25.5 (2018): 392...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating...are needed to deliver all the protein and RNA components, and several sgRNAs must be screened to identify...eBook Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

...target provided that (1) the sequence is unique compared to the rest of the genome and (2) the sequence...for the canonical S. pyogenes PAM sequence NGG compared to wild-type SpCas9, which also displays some ...CjCas9) is even smaller than SaCas9 and is also compatible with AAV delivery. NmCas9, another small ortholog...(2018) Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 ....

...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate...

... and the complexity of effectively deploying them in the lab somewhat mirrors the complexity of understanding.... Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both to..., and jGCaMP8 viral preps distributed per year compared to other AAVs preps from 2016–2023. B) Inset showing...

...virus as self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand...-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing, and ...virus’s most abundant proteins and a critical component for viral genome packaging. Addgene recommends...