Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 492 results for: ara-2

Showing: 181 - 200 of 492 results

  1. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ... However, if you have a larger knock-in (200 bp – 2 kb), a dsDNA donor will need to be used due to synthesis...accidentally change your protein of interest.   Fig. 2: Stop re-cutting with PAM and guide RNA disrupting... the other repair mechanisms Mammalian cells have 2 primary repair mechanisms besides HR - Non-homologous.... Com., 8(2024), (2017). 10.1038/s41467-017-01836-2 Charpentier, M., Khedher, A. H. Y., Menerot, S., Brion...recognition sequence in the donor DNA, you can guarantee there won’t be re-targeting after HDR. If your...

  2. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important ...1996 Apr 12;272(5259):263-7. PubMed PMID: 8602510. 2. A third-generation lentivirus vector with a conditional...CH, Humeau LM, Rebello T. J Gene Med. 2013 Feb;15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment... Banasik MB, McCray PB Jr. Gene Ther. 2010 Feb;17(2):150-7. PubMed PMID: 19847206. Additional Resources...Mulder GJ, Polack A, Bank A, Soulier J, Larghero J, Kabbara N, Dalle B, Gourmel B, Socie G, Chrétien S, Cartier...

  3. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ... to DFHBI in the presence of that metabolite (Fig.2). Spinach eventually evolved into Spinach2, a more...PMID: 25713381. PubMed Central PMCID: PMC4364232. 2. Baker, Monya. “RNA imaging in situ”. Nature Methods...emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067. PubMed Central PMCID: ...biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February 2008. 11. Svensen, Nina and Jaffrey...would bring the relevant ends together rather than separate them (Fig. 1b). One can also make “signal-off”...

  4. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...droplet in each well (Figure 2). A clean ddPCR should have a clear separation between positive (blue) and... to 2 minutes and the denaturation time to 1 minute has been shown to increase droplet separation (Witte... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...getting a clean separation between positive and negative droplets with these parameters, here are a few...Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the ... for ddPCR set-up. Prepare the master mix in a separate area from where you prepare your sample dilutions...your PCR For a good starting place on what PCR parameters to use, see Lock’s seminal paper on AAV titration...

  5. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...3 plates at different densities for selection). A 2% agarose solution is then melted (and thereby sterilized...96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with different levels of ...of agarose, which is then overlaid with the firmer 2% agarose gel solution discussed above for picking....concentrations of salts and glucose. If we circled well-separated clones containing more than 50 cells (visible ... should contain at least 50 cells and be well-separated. If the growth rate/transfection efficiency is... with this!) Make sure your colonies are well separated. For this, you need to have some idea of the rate...individual cell clones with different expression characteristics from a population of transfected clones. The...

  6. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the importance of... the open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...optimized Sep-OTSλ and can recommend its use (6).  2. C321, a specialized bacterial host strain for pSer-incorporation...biology4.5 (2002): E127-E130. PubMed PMID: 11988757. 2. Hornbeck, Peter V., et al. "PhosphoSitePlus, 2014...phosphorylated intermediate of a protein for characterization. However, phosphoproteins are often low abundance...strategies; expression should be optimized for parameters such as induction temperature and length of induction...

  7. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

    Type
    Blog Post
    ...mammal called alpaca could neutralize the SARS-CoV-2 virus. But while the early results were promising,...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a specific...Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert’s suitability and availability...diseases, so we were left with no choice but to quarantine in our houses, feeling helpless about the situation...

  8. Antibodies 101: ChIP

    Type
    Blog Post
    ...to (1) run a DNA gel to confirm fragment size and (2)  incubate with RNAse and Proteinase K to purify the...proteins of interest to any DNA they were bound to (2) fragmented the DNA (3) used antibodies to specifically...adjustments, to get everything just right!  Fig. 2: Goldilocks is actually celebrating a successful ChIP...remain bound to each other until you are ready to separate them. Most proteins are not likely to remain attached...

  9. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...multiple separate wells (within the same plate) with purified plasmid from the same aliquot/preparation. The...co-transfected with the test plasmid or transfected into a separate well of your cells. Regardless, the amount of ...should be consistent over time and from different preparations of the purified test plasmid expressing the ...Using our example above, we would test different preparations of Plasmid A in different aliquots of cells ...

  10. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ... Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin C, et al. An erythroid...with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore... inferred based on correlative biochemical characteristics and ectopic reporter assays. We have been interested..., identifying these deletions, and finally characterization. Read and download the full protocol from ...Dass A, Yien YY, Chung J, Masuda T, et al. Characterization of genomic deletion efficiency mediated by...

  11. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ... is also gaining a lot of new traction lately (1, 2), depends on knowledge of the structure(s) a given...per plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now...microbiology 23.4 (2015): 185-191. PubMed PMID: 25708933. 2. Cooper, Callum J., Mohammadali Khan Mirzaei, and ... biology 34 (2015): 69-77. PubMed PMID: 26275959. 2. Weigele, Peter, and Elisabeth A. Raleigh. "Biosynthesis... to be explored and exploited. With better characterization, these proteins could function as surrogate...surrogate antibodies for identifying and characterizing sugars and sugar-containing molecules in molecular biology...colonies of interest, sequence the inserted DNA and characterize your new putative host cell-binding proteins...

  12. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    .../10.1038/s41587-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, X., van der Weide, R. H.... flanking sequences to their input string (Figure 2). This string can then be used as input for the prediction...top 5) and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with pridict.it...we can evaluate tens of thousands of designs in parallel. To do this, we performed high-throughput ‘self-targeting...the TRIP (Thousands of Reporters Integrated in Parallel; Akhtar et al., 2013; Schep et al., 2021) method...Assayed by Thousands of Reporters Integrated in Parallel. Cell, 154(4), 914–927. https://doi.org/10.1016...., Benvenuto, E., Schmidheini, L., Schep, R., Damodharan, T., Balázs, Z., Janjuha, S., Ioannidi, E. I....

  13. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...vectors to answer this question, mapping and characterizing the activity of the DA neurons of the VTA. ...brain area, demonstrating that it is possible to separate neuronal inputs to induce aversion- or reward-...PMC4971574. Hooper, Andrew, and Jamie Maguire. "Characterization of a novel subtype of hippocampal interneurons...

  14. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ... delivered separately). Figure 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-...RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...mGold2t (Lee & Lai, et al., 2025). The team characterized and deposited a variety of constructs for visualizing...TnpB (named ISYmu1) to the popular plant model Arabidopsis (Weiss et al., 2025). By co-delivering TRV1 with...editor for transgene-free germline editing in Arabidopsis. Nature Plants, 11(5), 967–976. https://doi.org...

  15. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...its abdomen to produce yellow-green light (Figure 2). Despite the title of this post, this reaction is...; no chemical reaction takes place. Figure 2: Simplified chemical reaction catalyzed by luciferase... has a detailed list of luciferase options. Table 2: Properties of luciferase enzymes Name Natural ...interaction occurs, the luciferase emits its characteristic light. However, if a protein interaction brings...

  16. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...promoter and terminator flanking the CDS, and a Level 2 multigene vector with two other independently regulated...the sequence you are trying to clone. Figure 2: MoClo assemblies depend on compatibility of four-...sequentially for your Level 0, Level 1, and Level 2 assemblies using the plasmid sequence files provided...after each transformation. This could be as few as 1–2 colonies if you’re an experienced MoClo user, but ...MoClo kit through their online catalog pages. We guarantee this will save you a lot of headaches down the...

  17. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources... a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https...transfection). This differs from typical insect cell preparations, which we talk about below. Processing The viral...Plasmids and Resources References Shaw, G., Morse, S., Ararat, M., & Graham, F. L. (2002). Preferential transformation...

  18. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures...similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes it more ...retain their ability to be detected after sample preparation better than standard proteins tags like GFP. ...smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes for light and ...

  19. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...inherent brightness and spectral characteristics of the moxFPs are comparable to the non-optimized parental...protein activity. We encourage investigators to characterize FP-fusion proteins functionally relative to ...

  20. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    ...revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate and redifferentiate... more common delivery methods include retrovirus [2], lentivirus [4], adenovirus [5], Sendai virus [6]...2006. 441(7097): p. 1061-7. PubMed PMID: 16810240. 2. Takahashi, K., et al., Induction of pluripotent stem... Pluripotent Stem Cells. Cell Stem Cell, 2015. 17(2): p. 178-94. PubMed PMID: 26189426. 15. Si-Tayeb, ...induced pluripotent stem cells. Stem Cell Rev, 2015. 11(2): p. 242-53. PubMed PMID: 25578634. PubMed Central... to regeneration. Nat Rev Mol Cell Biol, 2011. 12(2): p. 79-89. PubMed PMID: 21252997. 37. Merrell, A....adapted from Zhou and Melton, 2008, [43]): Characteristic Reprogramming+ Transdifferentiation in vivo...
Showing: 181 - 200 of 492 results