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  1. Plasmids 101: Modular Cloning Applications and Kits

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    Blog Post
    ...Nicotiana Benthamiana and 12× Arabidopsis Mutants.” The Plant Journal, vol. 106, no. 1, Wiley, 25 Mar. 2021, ...within the mitochondrial matrix.     Figure 1: A chart with MoClo kit recommendations for different...Traditional Biology.” ACS Synthetic Biology, vol. 5, no. 1, American Chemical Society (ACS), 4 Nov. 2015, pp...., 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. ...Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer Science and Business Media LLC, 12 July ...
  2. Hot Plasmids: Summer 2024

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    Blog Post
    ...10.1038/s41551-024-01227-1. doi:  https://doi.org/10.1038/s41551-024-01227-1   CHARM: A compact epigenetic...spacer proteins ending with a GFP nanobody (Figure 1). This enrichment streamlines the purification and...highly-visible beads on the cryo-EM grid.     Figure 1: MagIC-cryo-EM for sample enrichment and structure...single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking...allowed the authors to determine structures at comparable or higher resolution than using more complex ...
  3. Choosing the B(right)est Fluorescent Protein: Photostability

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    ...photostability of the variants is comparable (lower panel of figure 1). To avoid changes in excitation ...various cyan fluorescent proteins is depicted in figure 1. Of note, the photobleaching rate does not depend ...to be used with the fluorescent protein. In figure 1 we provide an example of such a comparison between...
  4. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

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    ...separation is maintained by 2 sets of interactions : 1. Interactions between the non-target strand and a ...theoretically decrease off-target cutting. Figure 1: Strand separation and target DNA binding to the Cas9... improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered ...M694A/Q695A/H698A). This new variant displays comparable on-target activity and similar or decreased off-target...
  5. Using AAV for Neuronal Tracing

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    ...but also others including pseudorabies virus type 1, vesicular stomatitis virus (VSV; Beier et al., 2011...Skeletal Muscle AAV1, AAV6, AAV7, AAV8, AAV9 Table 1: Common AAV Serotypes and their target tissues (adapted... serotypes capable of entering neurons (see table 1)(Choi et al., 2005; Taymans et al., 2007; Howard et...efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One 8:e76310...Wolfe, J.H. (2014). Adeno-associated virus serotypes 1, 8 and 9 share conserved mechanisms for anterograde...,9 in rat neurons and glia in vitro. Virology 372(1), 24–34. PubMed PMID: 18035387. PubMed Central PMCID...transduction efficiency of recombinant AAV serotypes 1, 2, 5 and 8 in the rat nigrostriatal system. J Neurochem...
  6. Progress Towards a PAM-Free CRISPR

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    ...nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology.     Fig. 1: SpCas9 has sparse PAM coverage in the human genome...PAMs and is capable of editing at frequencies comparable to SpCas9. SpRY can also edit NYN PAMs, but with...
  7. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

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    ...effective recovery of knock-ins, but did result in 1-2 kilobases of additional sequence being introduced...facilitating recovery of edited animals. Figure 1: In the background are 504 restriction digests of ...selection for repair of a temperature-sensitive pha-1 mutation significantly enriches for knock-in of 2x... 3xFLAG epitopes into other, non-linked loci; pha-1(e2123) mutant worms are perfectly viable at 15 ºC,... repair pathway. Reagents required to perform pha-1 co-conversion are available through Addgene. In 2019...brilliantly and irreverently illustrated in the parable of the Geneticist vs the Biochemist, we geneticists...
  8. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    ...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...and spectral characteristics of the moxFPs are comparable to the non-optimized parental proteins. Our recent...
  9. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

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    ...restriction digests are comprised of 2 separate steps: 1) incubating your DNA with restriction enzymes which...The resulting gel image includes a 1kb ladder (lane 1) that has bands ranging from about 500bp to 10kb, ...will allow you to approximate the DNA mass of comparably intense samples of similar size. Run the gel ...
  10. Plasmids 101: Repressible Promoters

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    ...Bacillus. Appl Microbiol Biotechnol. 2019 Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018...and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55. PubMed PMID: 22205518. PubMed Central...Methods Mol Biol. 2016;1478:53-78. doi: 10.1007/978-1-4939-6371-3_3. PMID: 27730575; PMCID: PMC5270762. ... and lac promoters.” Proc Natl Acad Sci U S A. 80(1) (1983): 21-5. PubMed PMID: 6337371. PubMed Central...Tet-Off systems: second generation. https://www.takarabio.com/products/gene-function/tet-inducible-expression-systems...
  11. Hot Plasmids: Fall 2024

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    ...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9...matured considerably faster, with expression levels comparable to GFP during early embryogenesis (Figure 3)....
  12. Hot Plasmids - May 2022

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    ... Zebrafish) (Xiong, et al., 2021).    Fig. 1: Schematic showing the workflow for the BLITZ system...Biol. 2022. https://doi.org/10.1038/s41556-021-00836-1 Visual barcodes for live cell clonal multiplexing...SSAP system reports an efficiency of up to 20%, comparable to commonly used CRISPR/Cas9 methods for large...
  13. CRISPRainbow and Genome Visualization

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    Blog Post
    ...all the colors of the CRISPRainbow. Figure 1: CRISPRainbow was used to track the localization of...is available as a kit from Addgene.   References 1. Chen, Baohui, et al. “Dynamic Imaging of Genomic ... labeling efficiency was similar to that of a comparable FISH method. Importantly, they did not observe...
  14. Plasmids 101: The Promoter Region – Let's Go!

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    ...mammalian expression from human elongation factor 1 alpha Constitutive  Tends to give consistent expression...gene. araBAD General expression Promoter of the arabinose metabolic operon Inducible by arabinose and repressed...
  15. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

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    ...native peptide bond at the split site. Figure 1: The base editor is split into two plasmids and co-transfected... was also between 1013 - 1014 vg/kg, a dosage comparable to those currently used in human gene therapy...phytochrome-like fusion proteins from the plant Arabidopsis thaliana. In the presence of red light (650 nm...
  16. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

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    ...editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes...dCas to the genomic DNA of interest  Figure 1. When ABEs are in action they bind DNA so that the...found that the smaller version, ABE8.x-m, showed comparable editing behavior to ABE8.x-d. Overall, ABE8s ...
  17. Deep Dive: Statistical Tests (Comparisons)

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    ...assumes your data follows. Fig. 1: Examples of comparable groups with and without equal levels of... closely data points fall to each other (see Fig. 1). Alpha values – numerical risk allowance in a ...values from smallest to largest and ranking them (1 being the smallest value). It calculates the cut off...
  18. Viral Vectors 101: Voltage Indicators

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    ...depths (1 mm), important for certain applications such as in vivo tissue imaging.   Figure 1: Ace-mScarlet...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and is significantly...
  19. Plasmids 101: 5 factors to help you choose the right cloning method

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    ...your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly simplified...actually doing the experiments are also generally comparable so the best way to compare costs is to look at...least 50 cloning reactions. The ligase cost is comparable to that of the restriction enzymes and overall...
Showing: 41 - 60 of 83 results