...serotype of the packaged DNA and look for potential contaminants. The analysis is a 2-step process using Geneious...total reads are most likely to correspond to the contaminant. Each contig is then BLASTed against the NCBI...contaminated. For AAV-68544, the best match for the contaminant is Addgene plasmid #44362. Method validation,... were easily able to identify AAV-44632, the contaminant we mixed into the original AAV sample at 5%. ...software looks for and quantifies the presence of recombinants in recombinase-dependent sequences. Recombinase-dependent...powerful tool that can be used to identify DNA contaminants in AAV preparations and provide information ...Encapsidation of Prokaryotic Sequences during Recombinant Adeno-Associated Virus Production and Their in...

...to identify the critical NANOG binding sites: 1. Find Enhancers that have NANOG binding sites 2. Design...ChIP-seq experiment (think “GPS for proteins”) for NANOG, an essential transcription factor in embryonic ... just over 13,000 significant binding peaks for NANOG in the human genome. But not all of those 13,000...Design a CRISPR screen to target and disrupt the NANOG enhancers 3. CRISPR out the enhancers from step 2. ... ESCs die or differentiate Step 3 reveals which NANOG-related genes are critical to stem cell survival...CRISPR in the next section. In order to denote the NANOG enhancers from step 1 with genome math, we’ll need...shorthand from the field of epigenomics: H3 – one of NANOG’s associated histone proteins K4 and K27 – locations...

... GFP-binding nanobodies, the project really took off. Unlike most antibodies, nanobodies, which occur ...inside cells. Nanobodies binding GFP were designed in 2009. Subsequent work showed that nanobodies could be...and money needed to develop new lines. Using nanobodies to create GFP scaffolds for transcriptional activation...  References 1.Tang, Jonathan C.Y., et al. “A nanobody-based system using fluorescent proteins as scaffolds...Modulation of protein properties in living cells using nanobodies.” Nat. Struct. Mol. Biol. 17 (2010): 133–138....Fluorescent fusion protein knockout mediated by anti-GFP nanobody.” Nat. Struct. Mol. Biol. 19 (2012), 117–121. ...

... Encoded Nanothermometer. Nano Lett. 2022 Jul 27;22(14):5698-5707. doi: 10.1021/acs.nanolett.2c00608. ... size (~1500 bp) makes iGECInano suitable for AAV packaging.    Find iGECInano plasmids here!   Matlashov....13.503850; doi: 10.1101/2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent...using small monomeric NIR fluorescent proteins (miRFP670nano and miRFP720) and the Ca2+-sensing domain from...from Twitch-2B (Matlashov et al. 2022). iGECInano displays better response kinetics and photochemical ...

...editing CHARM: A compact epigenetic silencer Recombinant antibodies for marking astrocytes MagIC-Cryo-EM... linkers and spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines the purification...alpha helix) and target capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP...spacers could enable many future variations using nanobodies or scFvs for other targets or could be used for...https://doi.org/10.1016/j.cell.2021.03.025.    Recombinant antibodies for marking cell types in the nervous...s Neuroscience Antibody Collection to find a recombinant anti-GFAP [N206A/8R] antibody shared by James...other cell types in the nervous system!  Find recombinant anti-GFAP [N206A/8R] antibody here!   ...

...confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first...outgrowth of bacteria. Selection and confirmation of recombinant clones.    Figure 2: Overview of using...replacements and tags.  The typical frequency of recombinants is 1 positive clone out of 104 to 105 colonies...easy to remove the recombineering system once recombinant clones are generated.  A simple way to do this...integrates, drug selection is not required for maintenance. A temperature shift to 42˚C not only allows ... Court DL. Enhanced levels of λ Red-mediated recombinants in mismatch repair mutants. Proceedings of the.... 2. Daiguan Yu, Hilary M. Ellis, E-Chiang Lee, Nancy A. Jenkins, Neal G. Copeland, and Donald L. Court...

...second day of Antibody, Addgene gave to me…  A recombinant anti-GFP!    🎶On the third day of Antibody, ...fourth day of Antibody, Addgene gave to me…  A recombinant Anti-Arl13b!    🎶On the fifth day of Antibody...sixth day of Antibody, Addgene gave to me… A recombinant Anti-PSD-95!    🎶On the seventh day of Antibody...eighth day of Antibody, Addgene gave to me…  A recombinant Anti-DYKDDDDK!   🎶On the ninth day of Antibody...tenth day of Antibody, Addgene gave to me…  A recombinant Anti-APP!   🎶On the eleventh day of Antibody... of Antibody, Addgene gave to me…  A growing recombinant antibody collection!     ...

...Bacterial expression vector for His8-SUMO tagged recombinant protein pDest-Sumo - For prokaryotic expression...

...primate brains Endothelial-specific AAV delivery Recombinant anti-FLAG tag antibodies Peptide-assisted genome...throughout the non-human primate brain. Nature Nanotechnology, 18, 1241–1251 (2023). https://doi.org/10.1038....1038/s41467-023-38582-7. PMID: 37291094.   Recombinant anti-FLAG tag antibodies By Ashley Waldron Of...versatile. Whether you plan to purify or image a recombinant protein, the FLAG tag has proven itself a good...did you know that we distribute ready-to-use recombinant antibodies as well? Our collection includes a...available in trial sizes. Addgene recommends these recombinant antibodies for use in western blots, though the... variety of applications. Find anti-DYKDDDDK recombinant antibodies here!...

...nature can inform us on how to create better recombinant vectors. In this blog post, we’ll define plasmid...harness those features to create broad host range recombinant vectors.  A plasmid’s host range is determined...phylogenetic groups (Jain and Srivastava 2013).  Recombinant vectors that are made from broad host range plasmids...host-independent   Building broad host range recombinant vectors Replicons, which consist of a plasmid...broad host range plasmids can be used to create recombinant vectors that can replicate in different bacteria...to incorporating broad host range replicons, recombinant vectors can be modified further to increase the...cloning sites (Keen 1988).  Find broad host range recombinant vectors at Addgene!   References and resources...

... of 1:1 (Zeltner et al., 2010). However, for recombinant AAV2, the same study reported a physical-to-infectious...2010), while another reported that only 20% of recombinant AAV2 particles were full, compared to 50% of ...David Bell and Svetla Stoilova-McPhie, Center for Nanoscale Systems, Harvard University.    Which titer...annealing can be affected by the presence of protein contaminants, which is a factor in AAV titration since the...packaging of genomes can limit production of recombinant AAV-2." Gene therapy 6.7 (1999). PubMed PMID:... Lock, Martin, et al. "Characterization of a recombinant adeno-associated virus type 2 Reference Standard...wild-type AAV as benchmark for infectivity of recombinant AAV vectors." Gene therapy 17.7 (2010): 872-879...

...components into plants. Nanoparticles (carbon nanotubes, Kwak et al., 2019), DNA nanostructures (Zhang et al., ...chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol 14:447–455 . https://doi.org/10.1038...Hu P, Giraldo JP (2020) Targeted delivery of nanomaterials with chemical cargoes in plants enabled by a... Cunningham FJ, Fan C, Landry MP (2019) DNA nanostructures coordinate gene silencing in mature plants....

...few minutes of exposure to a pathogen and is the dominant defense strategy during the first several days...necrosis factor (TNF) family of proteins are predominantly expressed by immune cells and help to regulate...

...expressed in frame on the N- or C-terminus of a recombinant protein of interest. The tag was developed as...wash steps removes excess proteins and other contaminants. The protein of interest is then eluted using...wash buffer should be high enough to disrupt contaminant-metal ion binding without eluting the target ...more stringent wash conditions and remove more contaminants without losing your protein of interest. But...Genetic Approach to Facilitate Purification of Recombinant Proteins with a Novel Metal Chelate Adsorbent...native histidine-tagged protein expressed by recombinant vaccinia virus. Proc Natl Acad Sci U S A. 1991...

...expressing the recombinant plasmid to propagate. Any cells that contained non-recombinant vectors (re-ligated...ccd operon, which is responsible for plasmid maintenance during cell division. ccdB codes for the toxic...the number of clones that do not contain the recombinant plasmid and therefore makes the cloning process... but should never be used when selecting for recombinant plasmids. Check out our blog post on Common Lab...References 1. Bernard, P. “Positive Selection of Recombinant DNA by CcdB.” Biotechniques. 1996 Aug;21(2):320...

...more complex and the presence of helper virus contaminants have slowed the use of this system. New and ...detailed instructions on how to generate the final recombinant adenoviral plasmid construct, please see Addgene... references 1, 2.*Pro-Tip*: once the correct recombinant pAdV plasmid is identified, re-transform into...2-3 weeks) - Here you’ll produce the primary recombinant adenoviral stock (rAdV-S). HEK293 cells are transfected..., et al. "A protocol for rapid generation of recombinant adenoviruses using the AdEasy system." Nature...2009): 547. PubMed PMID: 19373227. 4. Mittereder, Nanette, Keith L. March, and Bruce C. Trapnell. "Evaluation...

...escape in SARS-CoV-2 Recombinant antibodies for aminoacyl tRNA synthetases Nanocompartments for yeast bioproduction...Validated recombinant antibody toolbox for aminoacyl tRNA synthetases By Kate Harten Recombinant antibodies...Clouse Claudia Vickers’ lab has created a novel nanocompartment for transporting proteins in yeast by using...virus-like particles (MPyV VLPs ). This artificial nanoparticle can protect proteins from unfavorable interactions...

...it’s possible to generate non-phosphorylated recombinant protein and perform in vitro kinase reactions...establishment of a system in which purified, recombinant protein could be produced with site-specific ...incorporation of phosphoserine directly into recombinant proteins (5). Optimization of this system defined...to effectively incorporate phosphoserine into recombinant proteins, such as the kinase MEK1 (5). MEK1 is...advantages of bona fide pSer-incorporation into recombinant proteins as an alternative to phosphomimetic ...Rinehart Lab reagents for improved expression of recombinant phosphoproteins’ available on the Addgene website...

... a set of techniques that are used to insert recombinant DNA from prokaryotes or eukaryotes into a synthetic...based approaches can require several hundreds of nanograms or micrograms of starting material; Gibson assembly...(genomic DNA and RNA), proteins and chemical contaminants that have been used during the purification ...scientists an indication of purity versus protein contaminants. The 260 and 230 nm ratios should fall ~2 - ... being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely endotoxin...

...promoter, AOX1, was co-opted for the expression of recombinant proteins by the pharmaceutical and biotech industries...efficient at protein secretion, making harvesting recombinant proteins a cinch. This is because it doesn’t ...secrete its own endogenous proteins, making the recombinant protein its major secretion. Beyond sheer protein... good for post-translational modification of recombinant protein products, specifically glycosylation....phaffii, relative to S. cerevisiae, is that it predominantly uses non-homologous end joining (NHEJ) to perform...