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We narrowed to 771 results for: INA

Showing: 121 - 130 of 771 results

  1. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    Published
    March 17, 2020, 1:32 p.m.
    ... TadA-7.10 version with a faster deamination rate so that deamination can happen before Cas homologs let...fused to a DNA-modifying enzyme, in this case a deaminase. Base changes from C•G-to-T•A are mediated by ... modifying enzyme to the target site, and the deaminase induces the DNA base change without a DNA double-strand... is ABE7.10 which contains the deoxyadenosine deaminase, TadA-7.10 (Gaudelli et al., 2017). The idea was...stop codons within T7 RNAP. A phage encoding the deaminase TadA-7.10 A mutagenesis plasmid controlling the...much higher base editing activity compared to the original TadA-7.10 enzyme. After testing numerous TadA ...10.1038/nature24644 Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW,...

  2. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    Published
    Jan. 30, 2020, 2:15 p.m.
    ... signals from time to time. False positives in original studies and false negatives in replications are...powered research designs. Unanticipated discoveries Finally, there are good reasons for obtaining a different...replication study. An important difference between the original setting and the new one, previously unaccounted...different handling can all be important.  If we eliminate the problems above: unclear methods, QRPs, and...when replications do not come out the same as an original research finding we can be more certain that some... papers. The precise timing, order, or every combination of materials and treatments is all too often ...reports, the gold standard for testing hypotheses Finally, Registered Reports combine all the best practices...

  3. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    Published
    June 25, 2020, 1:15 p.m.
    ...and Shannon Rinaldi in the Addgene lab! Read their Instagram profiles here (Kenkel, Rinaldi)! But leaving...might not be the right choice. I spoke with our Financial Operations Specialist, Lucy Hammer, about why ... have to go back to school right away. Shannon Rinaldi, Addgene’s Lab Manager, was taking classes at her...science, she stuck with it. Lab Manager Shannon Rinaldi values loving what she does. Her parents told her...response to this question came from Lucy Hammer, our Financial Operations Specialist. She said, “I’ve found the...came to Addgene with a goal to learn nonprofit finance, and while I've done that, my big takeaway has ...conversation master knows that although her focus is finance, it is just as important to bring love and fun ...

  4. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...and may increase the risk of mistakes or contamination. Finally, choice of concentration method depends...RCL). Replication competence can arise from recombination events during viral production and poses a biosafety...elements across multiple plasmids, the risk of recombination and consequently, the generation of RCLs is ...plasmid. In addition, the promoter of the 5’ long terminal repeat (LTR) is mutated in third generation systems...additional plasmids thereby increasing the number of recombination events that are required to generate RCL. To...volumes accordingly. While you may need to make some final adjustments, this is generally a good place to start... Keller B, Makalou N, Sutton RE. Systematic determination of the packaging limit of lentiviral vectors...

  5. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...list of Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism...depicted. (Bottom) Schematic depicting available destination vectors for various delivery methods, along with... with compatible inserts. All destination vectors contain an ampicillin resistance cassette and an origin...components you want, simply assemble them into a destination vector via Golden Gate cloning, perform an exonuclease...sequence for every individual assembly. What destination vectors are available? What you’re building ...Organism Delivery Type Destination Vector Mammalian Lentivirus pRDA_512 (RSV-driven; lentiGuide-based...type (lentivirus, AAV, transposon, Plasmid) Destination vector (dropdown options depending on above selections...

  6. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...Originally published Aug 3, 2017 and last updated Apr 6, 2021 by Will Arnold. Introducing NGS in 2017...plexWell technology to easily and quickly create Illumina sequencing libraries. From start to finish the...libraries are then QC’d, pooled, and prepared for Illumina sequencing on our newly donated MiSeq.  For Complete...electrophoretic gel, and qPCR quantification of the final libraries. A broader view of the whole plasmid To...plasmid.     Confirming tags and fusion proteins Finally, we confirm promoters, tags, fusion proteins, and... Analysis Platform to Assess the Identity of Recombinant Adeno-Associated Viral Preparations from Viral... also adds the necessary adapter sequences for Illumina sequencing as well as unique barcodes per sample...

  7. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...did not result in significant recombination. If you do see recombination, we recommend ensuring the cells...pooled libraries can lead to issues such as recombination and loss of plasmid diversity, so Addgene strives...strives to distribute aliquots of the original library whenever possible. In this blog, we’ll discuss ... lentiviral backbone might be more prone to recombination, while a library with a large backbone might...diversity of the library is maintained and reduce recombination during the amplification process. In liquid ...factor overexpression library) or is prone to recombination, we recommend amplification on solid media. ...may not have amplification protocols attached.  Final step: validation  Remember, there is no “one-size-fits-all...

  8. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ...protein tag which can be added to either the N or C terminal of a protein and can inducibly facilitate the ...concentration can yield intermediate protein abundance. Finally, some degron tags can do more than just degrade... the gene of interest and the degron domain is terminal. Between the tag elements (protease and degron...endogenous CRBN E3 ligase complex which will polyubiquitinate the fusion protein. This will target the protein...protein-of-interest and the CRBN E3 ligase complex leading to ubiquitination and proteasomal degradation. Adapted from Nabet...interacting ligands. After an E3 ligase facilitates ubiquitination of the Halo-tagged protein in question, it ...complex members, an E3 ubiquitin ligase complex. Finally, when auxin is added in the form of auxin hormone...

  9. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...unique repair templates. A final ligation reaction correctly assembles the final targeting vector (Fig. 1...need for PCR or cloning, as digestion of the destination vector with SapI yields 2 sites- the first site... the same unique SapI 5’ overhang, different combinations of connector and tag donor plasmids from the... a tissue specific manner, as well as 3-site destination vectors for inserting a tag at multiple target...the Hornung Lab, allows users to easily create C-terminal tag fusions of endogenous genes in human cells...plasmid containing the homology arms and an N- or C-terminal tag. The two PCR reactions are then mixed and ...expressing Cas9. This PCR toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag, YFP, Strep, TEV-V5...

  10. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...researchers test many different combinations of promoters, terminators, solubility and purification tags...connectors, cDNAs, and more to find the perfect combination. The EcoFlex Kit has a wide array of DNA parts...EcoFlex a great option for those interested in recombinant protein purification and metabolic engineering...expression, the system includes a toolkit to express recombinant proteins within the mitochondrial matrix.   ...specialized system with parts including site-specific recombinases, regulatory elements, developmental regulators... examine the effects of different promoters, terminators, enhancers, and cross-cassette interactions in...localization and transport studies in HEK cells, recombinant protein purification in bacterial cells, and ...
Showing: 121 - 130 of 771 results