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We narrowed to 46 results for: TRIP-1

Showing: 21 - 40 of 46 results
  1. Viral Vector Guides and Plasmids

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    ...integration. Introduction to Viral Vectors Figure 1: Wild-type virus propagation and use of viral plasmids...viral vectors, wild-type viral genomes have been stripped of any unnecessary accessory genes and divided...
  2. CRISPR Guide

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    ...changing the target sequence present in the gRNA. Figure 1: Overview of the basic CRISPR mechanism. CRISPR was...from Staphylococcus aureus ) has a coding sequence ~1 kb shorter than SpCas9 while retaining the same basic...lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included in the gRNA-containing...efficiency and fidelity. Nature Communications , 13 (1), 1425. PMID: 35301321 Edraki, A., Mir, A., Ibraheim...interrogation by SpRY-Cas9. Nature Communications , 15 (1), 3663. PMID: 38688943 Hsu, P. D., Scott, D. A., Weinstein...increase its specificity. Nature Communications , 9 (1). PMID: 30082838 Maddalo, D., Manchado, E., Concepcion...CRISPR-Cas9 with Bacteriophage Proteins. Cell , 168 (1–2), 150-158.e10. PMID: 28041849 Sakuma, T., Nishikawa...
  3. Viral Vectors 101: Producing Your rAAV

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    ...Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. https://doi.org/10.1089...plasmids, commonly called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps of...repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org/10.1089/hgtb.2011.034 Benyamini...in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https://doi.org/10.1186/s13024-020-00361-z ...
  4. Adenoviral Vector Production and Troubleshooting

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    ...repeats (ITR) present at both ends.   Figure 1: A) The structure of an adenovirus: the virion is ... 5 key steps: Construction of the rAdV plasmid (~1 week): The AdEasyTM system is the most popular method...Addgene’s Adenoviral Guide webpage, and references 1, 2.*Pro-Tip*: once the correct recombinant pAdV plasmid...transfected with the AdV plasmid construct from step #1 and allowed to stay in culture for up to 20 additional... it will result in very low titer. Amplification (1-2 weeks) - The rAdV-S is used to infect more HEK293...-4 rounds) at increasing scale over the course of 1-2 weeks. Each round of amplification should result...CsCl are commercially available. Titer calculation (1 or 10 days) Physical titer (particle count, rAdV ...
  5. Hot Plasmids February 2024

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    ...Biotechnology, 10.1038/s41587-023-01756-1. https://doi.org/10.1038/s41587-023-01756-1. PMID: 37095348. Breaking...incubation, even with multiplexed guides. Figure 1: A) Cell-penetrating Cas9, fused to HIV TAT, Myc and...expressing COX8a-mStayGold. Scale bars 10 μm (main), 1 μm (inset). Image reused from Ando et al. 2023, under... membrane-targeting applications. Nature Methods, 1–9. https://doi.org/10.1038/s41592-023-02085-6. PMID...StayGold fluorescent protein. Nature Biotechnology, 1–4. https://doi.org/10.1038/s41587-023-02018-w. PMID...broad tropism in primates. Nature Communications, 14(1), 3345. https://doi.org/10.1038/s41467-023-38582-7...generated a series of StayGold mutants and identified a triple mutant, Y187F, R144I and T155Q, which they named...
  6. Hot Plasmids - August 2020

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    ...membrane targetin sequence derived either from p63(1-29) and Lck. Image from Chertkova et al., 2020. ...Reports. 2020. https://doi.org/10.1038/s41598-020-57500-1 The CRISPR corner By Jennifer Tsang Here are a few... fluorescent proteins, they found that adding a triple nuclear localization signal or including fusions...CRISPRi-based synthetic oscillator, bistable network, stripe pattern-forming incoherent feed-forward loop. ...
  7. Plasmids 101: Secondary Nanobody Toolbox

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    ...incubation of 1° and 2° Must use different species and/or different IgG subclasses of 1° for multi-color...displacement 2° incubation can be skipped by pre-binding to 1° Monovalent Monoclonal Only anti- rabbit and -...dyes are also commercially available. References 1. Pleiner, T., Bates, M., & Görlich, D. (2017). A toolbox...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is...with the desired fluorophores. In figure 4B below, triple colocalization is possible even when staining cells...
  8. Using Phosphoserine to Study Protein Phosphorylation

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    ...translation (Figure 1): A tRNA that recognizes a specific ‘open’ codon and utilizes this triplet to incorporate...cellular proteins are modified through phosphorylation (1). However, the expansion of studies on protein phosphorylation... open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...modification allowed the deletion of release factor 1, the protein responsible for terminating translation...Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing... can follow her on Twitter @nieminm.   References 1. Cohen, Philip. "The origins of protein phosphorylation...
  9. RANbodies: Reporter Nanobody Fusions

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    ...learn more about each reporter or check out table 1 for a summary. HRP HRP tagged RANbodies don’t require...with a standard primary/secondary antibody. Table 1: Summary of the Key Features of HRP, Spaghetti Monster...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is...stained with two HRP RANbodies if the sample is stripped. HRP-RANbodies are, however, compatible with most...
  10. Neurodegeneration Research Collection

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    ...one of three different inherited genes: Presenilin 1, Presenilin 2, and APP. The majority (> 90%) of individuals...caused by abnormal expansion of a repeating CAG triplet series within the huntingtin (HTT) gene. An individual...
  11. Light Sheet Fluorescence Microscopy

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    ...microscopic resolution for studying colloidal gold (1).  The method was based on using a thin plane (sheet...Siedentopf, and R,. Zsigmondy.  (1902) Ann. Phys.  315, 1 – 39.
 A. H. Voie, D. H. Burns, F. A. Spelman. (1993...microscopy/en_de/products/imaging-systems/lightsheet-z-1.html ...along the illumination path. The shadows appear as stripes in the acquired image (5).  A way to get around...
  12. QC Sequencing Technologies at Addgene

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    ...and recorded as a DNA strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions...trace” file to determine the sequence. Figure 1: Overview of Sanger sequencing technology. Created...Technologies. Current Protocols in Molecular Biology, 122(1). https://doi.org/10.1002/cpmb.59  Wang, Y., Zhao,...or G) is supplied as a dideoxyribonucleoside triphosphate (ddNTP) so that after its addition, DNA synthesis...
  13. Plasmids 101: Protein tags

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    ...does not greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function... conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK 1 Affinity and Purification Good for antibody-based ...Unlike other epitope tags – which when doubled or tripled increase the tag size quickly – modifying the length...
  14. Viral Vectors 101: Transductions

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    ...cell type, and/or desired event.   Figure 1: Transduction to expression process. A viral particle...The triple T’s of biology are transformations, transfections, and transductions! In this blog we will...
  15. Antibodies 101: Multiplex Immunofluorescence

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    ...et al., 2019; McLaughlin, 2019).    Figure 1: Schematic of multiplex immunofluorescence approaches...). Springer New York. https://doi.org/10.1007/978-1-4939-8935-5_26 Jambor, H., Antonietti, A., Alicea,...by chemical treatment or photobleaching or by stripping out the antibodies, then start over with the next...
  16. Antibodies 101: The Basics of Western Blotting

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    ...to western blotting  In a western, proteins are: (1) separated by size, (2) transferred to a membrane,...  (3) detected using antibodies.   Figure 1: Overview of the Western blot process. The sample,... In addition, PVDF membranes can be repeatedly stripped of an antibody and reprobed with different antibodies...
  17. Visualizing Genomic Loci with CRISPR-Sirius

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    ...the guides are relatively unstable.   Figure 1: CRISPRainbow labeling. Two MS2, PP7, or boxB sequences...(by CRISPRainbow) to 3-fold with CRISPR-Sirius. Triple color detection With a third sgRNA containing alternating...
  18. 5 Great Apps for Lab Life

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    ...instead conveniently provide them on your smart phone. 1. Lab Archives by LabArchives The single, most important...can safely say that I still have the single and triple abbreviations of the amino acids memorized, but...
  19. Giving gRNAs a Facelift - Synthetic and Beyond

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    ...incorporated at the termini of the crRNA molecule (usually 1-3 of the first 3 bps are modified) for optimal stability... specifically ones which do not contain a 5’ triphosphate in the gRNA, reduce the innate immune response...
Showing: 21 - 40 of 46 results