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...but did not consider them for myself. I started picturing myself in a technical support role only when I...there is a second-line team that takes on cases requiring in-depth analysis, data review, in-house testing...are obtained. Or someone may heat an ELISA plate during incubation steps in order to enhance the OD signals...with other departments such as research and manufacturing, quality assurance, product management, marketing...

... not all, of these new experimental species. Discovering ways to culture, propagate, transform, store,...systems. How the EDGE Program promotes scientific sharing The EDGE Program is designed to expand the number..., in general, these labs all include materials sharing in their dissemination plans and in particular,...to mate with and if she decides to raise their offspring or leave them to their own devices. Chytrid fungi...

...for the entire translation process. For example, during high growth rate conditions, E. coli preferentially...biosynthetic enzymes may be preferentially charged during amino acid starvation thus resulting in higher ...getting your genes of interest codon optimized by ordering plasmids containing them from Addgene. If a plasmid...that will generally segregate with the cell pellet during purification procedures. Even if you produce a ...

...are occurring every year. At Addgene, we want to maintain the highest quality and accuracy during the ...analysis required every week. They are constantly exploring ways to enhance their processes, quality, and ...McQueary References and Resources References Niedringhaus, T. P., Milanova, D., Kerby, M. B., Snyder, ...

... Research Trust (RSRT) has an ambitious goal of curing Rett Syndrome and Addgene is thrilled to partner...methylated CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson & Bird, 2019) and...neurological deficits observed in Rett Syndrome. Restoring the MECP2 gene by excision of the Stop cassette...have shown exciting and promising results for repairing mutations in models of neurological disease. Klaus...

...biotin-streptavidin interaction (Figure 1a). This interaction brings the fluorophore in proximity to the target marker... used. These dyes are incorporated into the DNA during DNA synthesis and then diluted with subsequent ...negative live cells can still be easily gated on during the analysis. Conclusion Congratulations! You made...intracellular labeling protocol is necessary, requiring chemical treatment of the cells after surface ...

... allow cells to become persistently fluorescent during development. Stochastic multicolor labeling systems... tags instead of XFP to optimize it for tissue clearing, a method that makes tissues, such as the brain...visible. Chemical tags are more resistant to tissue clearing methods and can be labeled by synthetic fluorophores...Multicolor Brainbow imaging in zebrafish." Cold Spring Harbor Protocols 2011.1 (2011): pdb-prot5546. Richardson...

...scientific effort. As we like to say at Addgene: sharing speeds science. Here, we’d like to highlight the...Kleinstiver lab also worked on the opposite project: engineering Cas9 variants with a preference for specific ... L. A., & Zhang, F. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science (New York, ...Custom CRISPR-Cas9 PAM variants via scalable engineering and machine learning. Nature, 643(8071), 539–...

...tools this could be applied to. When it comes to altering gene expression, CRISPR technologies have gradually...-RNA (miRNA) Short hairpin RNA (shRNA) Short interfering RNA (siRNA) Piwi-interacting RNA (piRNA) miRNA..._module_attribute %}{% end_module_block %}   Interfering in the future While RNAi has retreated a bit ... the background, it remains a solid option for altering the expression of specific mRNAs. shRNAs are a...

...is a constantly evolving and demanding field, requiring a variety of biological and molecular tools. One... perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive research...be analyzed to understand human disease alleles during and after embryogenesis. Addgene has over 600 plasmids...organism: quick development, a large number of offspring (produces many seeds), self fertilizing, a small...

... the cut site were not efficiently incorporated during HDR. We decided to systematically characterize ...References 1. Cong, Le, et al. "Multiplex genome engineering using CRISPR/Cas systems." Science 339.6121 (...Mali, Prashant, et al. "RNA-guided human genome engineering via Cas9." Science 339.6121 (2013): 823-826. ...Fidelity Cas9 Variants How to Design Your gRNA Delivering CRISPR Ribonucleoproteins Resources on Addgene.org...

...), and in SNP analysis (to introduce naturally occuring SNPs in a plasmid context). The technique is also...sequence critical for Cas9 cleavage), thereby rendering the resulting construct resistant to Cas9 induced... successfully anneal to the plasmid of interest during the PCR reaction. Ideally, your primers should ...’ll want to steer clear of these strains when preparing the plasmid template for site directed mutagenesis...

...assembly work? The required homology between neighboring fragments can be created via PCR amplification...including regions of homology to the vector or neighboring fragment. Then you would amplify the fragments...required single-stranded annealing and priming of neighboring assembly fragments. Gibson assembly is commonly...modular overlap-directed assembly with linkers) which brings modularity to overlap-directed methods (Casini ...

...helpful from a biological standpoint, it makes delivering AAVs to the brain difficult. Traditionally, delivery...and/or its intermediate engineered precursor. Considering systemic capsids for your research? While systemic.../cJ and CBA/J mice. AAV1-X1 was developed by transferring the 7-mer peptide region from variable region...Fox AS. (2023). Advances in AAV technology for delivering genetically encoded cargo to the nonhuman primate...Bloom, J. M., Seed, C., & Deverman, B. E. (2019). Delivering genes across the blood-brain barrier: LY6A, a...AAV vectors to the central nervous system by engineering capsid–receptor interactions that enable crossing...

...recorded plasmid multimerization in the naturally occurring E. coli plasmid ColE1 (Goebel & Helinski, 1968...whole plasmid sequencing of our plasmid samples during our quality control process. Since MiSeq involves...D, Berg DE. pBR322-derived multicopy plasmids harboring large inserts are often dimers in Escherichia ...Guttman A, Thompson L. Vaccine Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med...

...2025). Nerve-to-cancer transfer of mitochondria during cancer metastasis. Nature, 644 252–262. https:/...microenvironment, immobilizing the components and altering the composition and dynamics. The researchers ... B) Constructs for tet-inducible CRISPRa-AAV engineering. Adapted from Nguyen et al. (2025) under CC BY...paper includes other great feats of materials engineering as well as extensive validation of the cell and...

...An off-target rate of 5% is far too high when considering human patients and the diseases associated with...off-target frequency is (more on this below). Lowering off-target events There are several strategies...compatible with the bulkiness that accompanies mispairing of bases. To reduce off-targets, you’ll want ...which rely on insertion of DNA by the NHEJ pathway during DNA repair of the break to allow for site-specific...

...the five is different. To identify significant pairings, additional tests, referred to as post-hoc analysis...temperature, humidity, sunlight exposure time, and watering volume on total crop production (though you would... error is the probability of a false positive occurring when many comparisons are performed. For example...drawbacks to these tests as they can be quite stringent if many samples are assessed. However, if many...

...minimize recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019)... showing the mechanism of spontaneous reversion during AAV plasmid amplification. On the left homologous...  Some classes of neurons transiently express TH during development or have low levels of TH mRNA without...time, frustration, and funds in the long-run by ensuring you are minimizing any off-target expression that...

...insertions were positive or negative selected for during the experiment (Sandoval-Villegas et al., 2021)...Targeting) system. INTEGRATE is based on a naturally occurring Tn7 transposon found in Vibrio cholerae that encodes...Sleeping Beauty, piggyBac and Tol2 for Genome Engineering. IJMS 22:5084. https://doi.org/10.3390/ijms22105084...about the INTEGRATE system for bacterial genome engineering  Read about the Sleeping Beauty DNA transposon...