We narrowed to 747 results for: cat.1

...wondering what her cat is thinking. Find her on Twitter @JessicaSacher.   References 1. Nobrega, Franklin...which is also gaining a lot of new traction lately (1, 2), depends on knowledge of the structure(s) a given...RBPs successfully identified using this method. Step 1: Prepare a random fragment library of phage DNA This...first step involves digesting phage genomic DNA into 1-3 kb (approximately gene-sized) fragments so that ...in the bulk of resultant fragments falling between 1 and 3 kb results in optimal likelihood of capturing...solution-soaked filter paper (colony-side up) and leave for 1 h at room temperature. After this, it is helpful to...proteins expressed by your library. After blocking for 1 h at room temperature with gentle shaking, gently ...

...first before larger fragments. Figure 1: The quantification of a RNA ladder using capillary electrophoresis...automated. Unlike gel electrophoresis, you only need 1-2 ul of sample and the run time is just a few minutes...see any signs of DNA in your final tube after purification. How do you know if you actually have DNA in...you choose could be based on your downstream application, time, and instrument availability. UV absorbance... at 230 nm. The A260/A280 ratio is used as an indicator of DNA purity. Ideally, this number should be ...like a PCR product). This method also gives an indicator of DNA or RNA contamination based on the presence... about quantifying DNA In choosing your DNA quantification method there’s many things to consider: cost...

...the journal-specific guidelines.  Peer-review step 1: The first read At the first read of the paper, I ...group of Prof. Mark Tester, where she focuses on (1) salt-induced changes in root-to-shoot ratio in Arabidopsis... to help decide whether work is suitable for publication in their journal. The early peer-review attempts...information that would prevent the study to be replicated or omission from the literature. I think that...Addgene.org Learn about depositing plasmids pre-publication Find molecular biology protocols Find eBooks ...

...General Design Figure 1: Two proteins are fused to the FP(11) and FP(1–10) fragments. When Protein...and 11 to yield large (1–10) and small (11) fragments. In several cases an FP(1–10) fragment may be compatible... to Plasmids) PI Blue EBFP2 EBFP2(1–10) and Capri(1–10) for use with GFP(11) Multiplexed labeling...Commun. 2017 Bo Huang mNeonGreen3 mNG3K(1–10) and mNG3A(1–10) for use with mNG2(11) Improved yellow-...sfCherry2 sfCherry2(1–10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1–10) Improved split...brings the pieces of the FP back together (Figure 1). This method is generally known as Bimolecular Fluorescence...even from a different base FP (for example, EBFP2(1–10) can be used with GFP(11)). Some of the articles...

... Antigen-1 (oriP-EBNA-1) system where each vector contains a viral origin of DNA replication, OriP and...pCXWB-EBNA-1 is used in the reprogramming cocktail. pCXWB-EBNA-1 lacks oriP and is unable to replicate in human...and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two sequences are necessary...expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein expression...pCXLE vectors listed below in Table 1 are available at Addgene. Table 1: List of pCXLE episomal plasmids,...sufficient for retention and replication of the vectors in human cells [8]. The EBNA-1 protein recognizes OriP...obtained and the subsequent reprogramming success [1]. Fibroblasts need to be cultured and expanded in ...

...several advantages over traditional vaccines, including 1) cellular responses in addition to antibody response...and immunogenicity of attenuated, nef-deleted HIV-1 strains in vivo. Retrovirology. 2007 Sep 23;4:66. ...their use in other clinical applications. In the last decade, viral applications in cancer treatment and ...s unpack the myriad number of emerging viral applications and give you some recommendations for putting...putting viruses to work. Viral applications Viruses work by infecting a host cell (the target of viral infection...expressed. It’s important to note that for all the applications we discuss, there is more than one type of virus...retrovirus, AAV, etc.). When you’ve decided on an application, our Viruses as Biological Tools blog can help...

... developed calcium indicators spanning nearly the entire visible spectrum (Figure 1). The GCaMP series...available in your microscope setup. Figure 1: Excitation (dotted) and emission (solid) spectra ...potential (AP) spiking events are indicated below the trace; asterisk indicates a single AP. B) Averaged fluorescence...past few decades, genetically encoded calcium indicators (GECIs) have become a vital tool in neuroscience...toolbox, our team recently developed a new calcium indicator, OCaMP. The challenge: Expanding the fluorescent... signals in the brain. For this to work, each indicator needs to emit a different color of light and be...gold standard for green fluorescence, while red indicators jRGECO1a and jRCaMP1a are widely used as well...

...one of three different inherited genes: Presenilin 1, Presenilin 2, and APP. The majority (> 90%) of individuals...loci seem to be associated with the disease and implicate a wide range of cellular processes. The vast majority... consists mainly of managing symptoms through medication and surgery. PD primarily involves the malfunction...available from Addgene, sortable by type of genetic modification and target species. You can also find our guide...synaptic transmission with improved glutamate indicators . (Aggarwal et al. bioRxiv. 2025) Monitor dopaminergic...window) A news website and information resource dedicated to helping researchers accelerate discovery and...Foundation (Link opens in a new window) A foundation dedicated to finding a cure for Parkinson's disease through...

...Addgene, we use seven different types of “plates” (Fig. 1). When you have to tell an engineer that you want ...”? We took steps towards solving this problem by 1) making sure we qualified these words when we discussed.... I needed to a) figure out the sources of miscommunication and b) find practical solutions.  My PhD ... for this task. Good scientists learn how to communicate their work differently based on their audience... family is going to vary widely. The way you communicate your science at a poster is going to vary depending...two main problems with Addgene’s cross team communication. Although these real world examples did not ... for the container to be used in cross-team communication A list of “AKA” names, or jargon terms, for ...

...    Figure 1: A 1 mg/mL control is diluted in a series ranging from 1:2 to 1:32. Each individual...trendline. R2 ranges from 0 to 1 with 1 being a perfect fit and 0 indicating that there is no linear relationship...example, Figure 1 depicts a 2-fold dilution series. Each step of the series dilutes by 1:2 for a series...series that ranges from 1:2–1:32. The specific dilution series used will depend on the expected concentration...individual step of the series is consistent at 1:2. When preparing a dilution series, use a new pipette tip for... be run in duplicate or triplicate. The closeness of the data points generated by replicate values provides... the standard dilution series in duplicate or triplicate. Replicates allows you to calculate the curve...

...Table 1 below outlines a few of the more common genetic changes found in E. coli strains. Table 1: Common...pooled library amplification. F' mcrA Δ(mcrBC-hsdRMS-mrr) recA1 endA1 gyrA96 gal-thi-1 supE44 λ-relA1 ...amber (UAG) stop codon by tyrosine insertion λ-thi-1 or thi1 Mutation in thiamine metabolism Requires exogenous... all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain Natural...plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF...) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109   General cloning and...strain for cloning repetitive DNA. endA1 glnV44 thi-1 relA1 gyrA96 recA1 mcrB+ Δ(lac-proAB) e14- [F' traD36...

...of 0.3, as recommended by the original publication. Pro-Tip: 1. Use multiple gRNAs to target different..., as indicated by the presence of a 428 bp digestion product (lane 2). The undigested (-, lane 1) DNA ...The entire process from start to finish took about 1 month. Cas9 delivery Overview: Before any editing...lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then used to generate a monoclonal...rest of the cells were frozen down. Figure 1: Cas9 expression in A549 cells transduced with lentiCas9...Western blotting is a loading control. Pro-Tips: 1. It’s all about the MOI. In retrospect I used a way... pool had wildly variable Cas9 expression (Figure 1). This is a problem because Cas9 expression affects...

...promoters. Gene Therapy, 28(1–2), 56–74. https://doi.org/10.1038/s41434-020-0169-1 Oh, J. Y., Han, J. H., ...AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), 1–41. https://doi.org/10.3390...constitutive promoters located in adeno-associated viral vectors. PLoS ONE, 9(8), 1–10. https://doi.org/...the selection process. Factors to think about are (1) your experimental conditions and measurements; (2...serotype, promoter, and titer.     Figure 1: Flow chart depicting a) steps for selecting serotypes...injection site than AAV2 (Watakabe et al., 2015) (Figure 1). You’ll need to consider if such properties could...

...chromosome, start, and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning of .... Table 1: An emerging family of "GPS for the genome" techniques Technique What It Locates ChIP-seq...clinical applications of CRISPR in the next section. In order to denote the NANOG enhancers from step 1 with...genetic elements like protein, RNA, and DNA (Table 1). As with GPS in the real world, coordinates alone...enhancers with the help of genome math. Figure 1: Genome math illustrated. Intersect (left) and set...histones with “GPS for proteins,” ChIP-seq from Table 1. Because of DNA’s 3D geometry and the chemical properties...experiment to identify the critical NANOG binding sites: 1. Find Enhancers that have NANOG binding sites 2. Design...

...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately... doing the purification and cross-adsorption in lab, consider a dilution range from 1:100–1:1,000. If ...recommended dilution and/or range (typically between 1:500 and 1:10,000 for primary antibodies). You’ll want to...from ascites fluid, try a dilution range from 1:1,000–1:100,000. Secondary antibodies For secondary antibodies...recommended dilution range is usually between 1:5,000 and 1:200,000. Check the manufacturer’s recommendations...antibodies that are not.   Table 1: Troubleshooting a western blot *indicates a diagnostic-only step Issue...reagent (usually an antibody).      Figure 1: A protein gradient. The density of the bands, which...

...recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three are required...that occur during DNA replication. There are two ways to avoid activating MMR: 1) use a strain of bacteria...Table 1: Overview of ds- and ss-DNA Uses in Recombineering Substrate Best Suited Application Recombination... interested in science communication and in vitro diagnostics.     References 1. Costantino N, Court DL...from the 5’ end and generate 2 possible products: 1) a partially dsDNA duplex with single-stranded 3’ ...recombineering with an ssDNA oligo substrate.   Figure 1: Components of the Lambda Red Recombineering system...and tags.  The typical frequency of recombinants is 1 positive clone out of 104 to 105 colonies. Pro Tip...

... assembly reaction** 1 5 min., 37 ºC (2.5 μl) 687 100% 274,200 2,742,000 1 60 min., 37 ºC (2.5 μl)...References 1. Vladimir, Potapov, et al. "Optimization of Golden Gate assembly through application of ligation...expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a mixture...representative transformation plates obtained from 1-, 12- and 24-fragment assemblies of the lacI/lacZ ...lacZ cassette, and illustrates how the volume of the 1 ml outgrowth spread on each transformation plate can...fidelity studies conducted by Potapov et al. at NEB (1, 2). While T4 DNA Ligase, the mainstay of most biotechnological...over 90% for 24-fragment assemblies (Figure 3, Table 1). These improvements (5- to 12-fold increases in the...

...   Class 1 versus class 2 There are two CRISPR classes: Class 1 and Class 2. Class 1 CRISPR systems...archaea, though Class 1 remains largely underutilized compared to Class 2. Class 1 Despite their incredible...optimal plasmids to harness the power of Class 1 systems. Class 1 CRISPR systems can be further broken down...and 17 variants. Figure 1: Overview of the CRISPR classification hierarchy. The hierarchy flows...effector complex after binding to a target. Class 1 is the most abundant of the two, comprising about ...incredible abundance, Class 1 CRISPR systems are less popular in the lab. This can be attributed to the multi-Cas...Type I CRISPR systems make up the majority of Class 1, and are the most common type of CRISPR overall. Type...

...depths (1 mm), important for certain applications such as in vivo tissue imaging.   Figure 1: Ace-mScarlet...Voltage indicators are voltmeters for cells! In this blog we will review how these indicators work, their...fluorescence or voltage indicators.   Viral Vectors and Voltage Indicators As previously mentioned...Enhanced genetically encoded voltage indicators advance their applications in neuroscience. Curr Opin Biomed...Cellular communication and function depend on potential energy. Cellular activities like action potentials...their general applications, and why viruses are essential to their use as biological tools. Why Use Voltage...Voltage Indicators? Electrical signals are integral to many facets of biology. For example, many heart...

...pipetting or irregular membrane transfer. Figure 1: A loading control is a type of positive control necessary...because the loading control band is weaker in condition 1 than condition 2, the normalized values for the two...to using loading controls accurately. Assumption #1: The loading control is not perturbed by the experiment...without your knowledge? If you can’t trust Assumption #1, any change in the normalized intensity of your POI...interest band is significantly lighter in “condition 1” than “condition 2.” By eye, the loading control appears... the protein abundance axis, both blue “condition 1” points (POI and loading control) are smaller than...assumptions discussed above into question. Assumption #1: The loading control is not perturbed by the experiment...