We narrowed to 718 results for: des.1

...transcription activation system co-opted from yeast (1). It is a Drosophila geneticist’s main workhorse to...he still reads “#” as a “pound symbol”. References 1. A. H. Brand, N. Perrimon, Targeted gene expression... A Fly Geneticist’s Swiss Army Knife, Genesis 34, 1–15 (2002).PubMed PMID: 12324939. 3. S. E. McGuire,...Versatile Toolkit for Gene Expression in Drosophila, 3, 1–10 (2015). PubMed PMID: 21356876. 5. J. Chow, Z. Marka...references and web links from my first post and start designing your first fly experiment! Many thanks to our...PMID: 28704652. 6. V. Rodríguez, D. Didiano, C. Desplan, Power tools for gene expression and clonal analysis...

...premature chain termination 1-10 ug/mL Zeocin Sh bla HeLa, NIH3T3, CHO, COS-1, 293HEK, Jurkat T cells Complexes...expressing the Epstein–Barr virus (EBV) nuclear antigen 1 (EBNA1) or the SV40 large-T antigen (293E or 293T ...Concentation*** Blasticidin bsd  HeLa, NIH3T3, CHO, COS-1, 293HEK Inhibits termination step of translation ...mammalian plasmid components, it may be useful to describe the means of introducing genetic material (such...the plasmid in the same way. Selection methods, described below, are typically employed when creating stable...

...how scientists connect on Twitter that I realized 1) a lot of scientists are using social media professionally... but my consistent writing for the Addgene blog (~1-2 blog posts per month) and use of Twitter and LinkedIn... have tips for using social media as a scientist. 1. "5 Ways Scientists SHOULD Be Using Twitter" podcast...of interest. Hopefully through these links, I’ve: 1) convinced you social media can positively impact ...’s social media toolbox. Besides advice for using Twitter, this post provides suggestions for using LinkedIn...use Twitter as a scientist. While this post is designed with beginners in mind, there’s some great reminders...sheet from Science Media Savvy This tip sheet provides advice for tweeting and blogging as a scientist...

...for more details on how they work!) Figure 1: Base transitions are edits between the purine bases...R-loop structure. In order to bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and...given the pithier name CGBE1, for “C → G Base Editor 1,” and had a C → G editing efficiency of up to 68%... the CGBE: DNA polymerase D2 (POLD2), exonuclease 1 (EXO1), and RNA binding motif protein X-linked (RBMX...excision repair proteins. Nature Communications, 12(1), 1384. https://doi.org/10.1038/s41467-021-21559-9...transversions in human cells. Nature Biotechnology, 39(1), 41–46. https://doi.org/10.1038/s41587-020-0609-x...machine learning methods. Nature Communications, 12(1), 4902. https://doi.org/10.1038/s41467-021-25217-y...

...Updated Jun 1, 2021 by Meghan Rego. Reproducible data are key to science, so scientists are used to repeating...the DNA packaged inside the viral particles (Figure 1). Briefly, packaged DNA is extracted from purified.... When contamination is suspected, we proceed to de novo assembly of the unmapped reads, employing our...transduced cells whenever possible. This post will describe our workflow for viral genome sequencing (VGS)...

...crevices proteins on the cell surface. Figure 1: Comparison of common antibody, HCab, and nanobody...Humana Press, pp 133–148. https://doi.org/10.1007/978-1-62703-293-3_10 Walter J et al. (2020) Synthetic nanobodies...https://doi.org/10.1101/2020.04.16.045419.  Wrapp D, De Vlieger D, Corbett KS, Torres GM, Wang N, Van Breedam...Zimmermann et al., 2020). This selection platform was designed so that any standard lab can quickly select for...strongly to your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the... randomized with a defined set of different trinucleotides (corresponding to different amino acids) in...for binding to the target protein. The protocol describes going through two rounds of phage display to increase...

...necessary.   Tips for barcoding and tracking samples 1. Start with the tube - Barcoded tubes come in many...all of these samples have the 2D barcodes mentioned above. These barcodes identify what type of sample is...tubes. 1D barcodes are better for tubes that are meant to be read one at a time. 1D barcodes can be purchased...the tubes’ physical location in the lab. These barcodes additionally allow us to store multiple types ...fresh media with antibiotics at each stage. The barcodes and LIMS help us track each sample, but we also... a petri dish plate that is also barcoded. The barcodes on both of these are scanned into our LIMS system...samples within your lab, we highly recommend using barcodes! Barcoded tubes are easily accessible and can ...

..., rather than a broadly  reactive secondary (Fig. 1). For example, an anti-mouse IgG2a secondary would...IgG2b > IgG1 (Manning 2012).      Figure 1: Broadly reactive versus subclass-specific antibodies...antibody binding and therefore the signal strength (Fig. 1). Monoclonal primaries If you do decide on a monoclonal...raised in a different species.      Table 1: Isotype considerations when choosing antibodies ...Elsevier. https://doi.org/10.1016/s0091-679x(08)60255-1  Manning, C. F., Bundros, A. M., & Trimmer, J. S. ... and to work in a variety of applications. The desired specificity of your secondary antibody depends ...

...journal publication describing the plasmids, or (for deposits submitted after July 1, 2025) after three... emailed submission) in the Deposit menu: Figure 1: Begin the deposit process by clicking "Submit Online...cases Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if your deposit falls into...Addgene?" or contact us at [email protected] or +1 (617)-ADDGENE (233-4363) with additional questions...strain. Please contact us at [email protected] or +1 (617)-ADDGENE (233-4363) if you’d like to distribute...page so that any data you enter is not lost.   Step 1: Sequences, Maps, and Files Please provide any full...a 1.5 mL microfuge tube at a concentration of 0.1–1 µg/µL (or bacterial streaks, if you prefer). If you...

...recently that desired genome-editing events by HDR can be generated more efficiently by (1) improving editing...specifically incorporate homo- or heterozygous mutations. 1. Increasing HDR accuracy with CRISPR/Cas-blocking ...targeted alleles when gene editing.   Figure 1: Techniques to improve editing efficiency.   But...provided by blocking mutations. Essentially, you:  Step 1 - Introduce the blocking mutation together with the...developing novel iPSC-based model systems.     References 1. Cong, Le, et al. "Multiplex genome engineering using...genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al, Mai et al) we were very excited...introduced. Although we were able to introduce the desired mutations in genes associated with dementia, such...

...Combinatorial Design for Synthetic and Traditional Biology.” ACS Synthetic Biology, vol. 5, no. 1, American...within the mitochondrial matrix.     Figure 1: A chart with MoClo kit recommendations for different..., vol. 4, no. 9, American Chemical Society (ACS), 1 May 2015, pp. 975–86. Crossref, https://doi.org/10.1021...Yeast Mitochondria.” BMC Biotechnology, vol. 17, no. 1, Springer Science and Business Media LLC, 13 Nov. ...Editing in Plants.” BMC Plant Biology, vol. 20, no. 1, Springer Science and Business Media LLC, 23 Apr. ...Arabidopsis Mutants.” The Plant Journal, vol. 106, no. 1, Wiley, 25 Mar. 2021, pp. 8–22. Crossref, https://...Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media LLC, 12 July ...

.... Level 1: Assembly of multiple Level 0 parts into TUs. Level 2: Assembly of multiple Level 1 TUs into...: promoter, gene, and terminator.     Fig 1. MoClo enables individual parts to be assembled into... peptide can easily be tested by generating Level 1 TUs with unique promoters, and the ideal phosphatase...cloning, or MoClo, is a systematic cloning strategy designed by synthetic biologists to perform large scale...specifically refers to the organizational schema described above, and not any particular cloning technique...example of an individual part, this category also includes promoters, UTRs, terminators, etc. Once selected...behave in a cell. For unpredictable cases, MoClo provides two answers: first, sets of parts with characterized...

... The three VP proteins assemble into capsid at a 1:1:10 ratio (VP1:VP2:VP3). There are eleven naturally...closer look at the rep and cap genes. Figure 1. The RepCap plasmid and its products. Created with...plasmid The packaging plasmid aka the RepCap plasmid encodes two genes: the replication (rep) gene and the capsid....com   Replication (rep) gene The rep gene encodes four proteins that are required for viral genome...cross-packaging.  Capsid (cap) gene The cap gene encodes three AAV viral capsid proteins VP1, VP2, and VP3...type tropism. Introducing changes and/or short peptides sequences into these variable loops is one method...method for changing a capsid’s tropism.  cap also encodes the Assembly-Activating Protein (AAP) and the recently...

...such as IL-8, IFNγ, and colony stimulating factor 1 (CSF-1). Additionally, tumor cells recruit regulatory...introduced the “Three Es of Immunoediting” nomenclature. 1. Eliminate - Latent tumors are recognized by effector...the tumor, activates hypoxia inducing factor (HIF-1), which promotes the production of VEGF to increase...This study used CRISPR gene editing to knockout PD-1 on immune cells harvested from patients and thereby...exploring new avenues in healthcare.   References 1. Vinay, Dass S., et al. "Immune evasion in cancer:...lymphoblastic leukaemia in children and young adults: a phase 1 dose-escalation trial." The Lancet 385.9967 (2015)... recognition are a few methods by which tumors sidestep immune surveillance: Suppressing Function Antigen...

...Table 1 has a detailed list of the bacterial species these plasmids are compatible with.  Table 1: Non-...reduced metabolic load (Meyer 2009). Figure 1: Idealized examples of a narrow host range plasmid...for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197. https://doi.org/10.1016/0378-1119(88)90117... N.J.), 765, 327–343. https://doi.org/10.1007/978-1-61779-197-0_19  Meyer, R. (2009). Replication and ... meliloti, Rhodobacter sphaeroides, R. capsulatus, Rhodopseudomonas spheroides, Salmonella spp., Serratia...Brevibacterium methylicum, Caulobacter crescentus, Desulfovibrio vulgaris, Erwinia carotovora, E. chrysanthemi...meliloti, R. leguminosarum by. viciae, Rhodobacter sphaeroides, Salmonella typhimurium, Vibrio cholerae, Xanthomonas...

...main takeaways from the paper for designing experiments with AAV: 1. Getting your viral vector into your... administration routes for neuroscience see Table 1 from the paper. 2. Manipulate your cell of interest...information from a region. In the example shown in Figure 1, injected retrograde AAVs at one location result in...AAV from our viral service can share experimental design and data. With these different approaches to open...

... the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the targeted gene have...through an experiment designed to knock out exon 2 of a given gene.   Figure 1: A knockout targeting... the Neomycin resistance (NeoR) gene between exon 1 and 3 of the targeted gene. Upon recombination, the...often use the Cre/Lox system described earlier. In this case, you design your targeting vector such that...replacing it or disrupting it with an piece of DNA designed in the lab. Specially constructed plasmids can...template with the endogenous gene. Here, we'll describe the techniques and the plasmids used to inactivate...inactivate specific genes in mammalian cells. Despite the popularity of CRISPR-based knockout/knock-in systems...

...choosing direction of a project. If you expect more 1:1 time with the advisor, you should choose accordingly...potential hire visits the lab for a full day, gives a 1 hour talk about his or her interests, and spends one-on-one...that supports both paths – you won't know in year 1 what you will want in year 6. Have lab alumni taken... a Good Mentor for Scientists". This post also includes a useful list of questions for a candidate to ...

...under a wide range of inducer concentrations (0.1-1% L-arabinose) without signs of toxicity or burden ...bacteria have colonized (Fig. 1c).     Fig. 1: Expressing bARGSer in E. coli. (a) Plasmid diagram...ultrasound contrast. In all the cell lines we tested, 1 µg/mL doxycycline was sufficient to saturate expression...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the ... tumors and tumor-homing bacteria. Nat Biotechnol 1–13 (2023) doi:10.1038/s41587-022-01581-y  ...to distinguish from background tissue unless destructive ultrasound imaging was used. We set out to develop...acoustic properties of these gas vesicles made non-destructive specific imaging in tissue difficult. The first-generation...

...applications. Figure 1: Schematic of an RNAi-inducing transgene. Typically, the desired target of RNAi is...dsRNA via an RNase III enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of...in the biogenesis of different small RNAs (Figure 1). These siRNAs can exert their gene silencing effect...modifications of complementary gene sequences (Figure 1). As a result, gene silencing can be easily induced...complementary to your gene target as inverted repeats (Figure 1). With this technique, multiple genes can be simultaneously...selection of plants undergoing RNAi, and LIIbeta F 1-2 RNAi, which allows assembly of intron-spliced hpRNAs...eukaryotic process where approximately 20-30 nucleotides of double-stranded RNA (dsRNA) results in downregulation...