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We narrowed to 400 results for: POR C

Showing: 101 - 120 of 400 results
  1. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ....1126/science.abb2920 Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH (2020) CRISPR...and require only two transposase proteins (TnsB and C). While requiring fewer components for RNA-guided ...Rubin BE, Diamond S, Cress BF, Crits-Christoph A,  He C , Xu M, Zhou Z, Smock DC, Tang K, Owens TK, Krishnappa...genomes engineering In the Sternberg group’s initial report of VchINT, three separate plasmids and multiple...simple to use, users should keep aware of a few important details. For transposons to be specifically recognized...
  2. Viral Production at Addgene

    Type
    Blog Post
    ...80 °C. Titer All titering is performed on lentiviral preparations that have been stored at -80 °C and ...Preparations are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or ...preparations are then aliquoted, frozen, and stored at -80 °C. Concentrated lentiviral preparations (“-LVC” catalog...expression and/or function. These data are sometimes reported or posted on the material page for the corresponding...recipient’s initial thaw will be accounted for in our reported titers. Lentiviral vectors are titered using a...
  3. Antibodies 101: Epitope Tags

    Type
    Blog Post
    ...Epitope tags are short peptides introduced at the N or C terminus of a protein that are bound by antibodies...enterokinase, which recognizes the 5 amino acids on the C terminal of the tag. Of the most common tags, FLAG...not at all. The introduction of a tag at the N or C terminus of a protein may disrupt the biological function...destabilization can be brought about by the tag so it’s important to validate tagged protein functionality (Arribere...
  4. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ... Boeckman, J. X., Manning, C. F., Nguyen, J. T., Bechtold, H., Dumitras, C., Gong, B., Nguyen, K., Van...-GABA-AR β1 (white) and anti-GABA-AR β3 (green). (C) Hippocampus labeled with anti-Kv2.1 (magenta), anti-AnkyrinG... Panels C1-C3 show magnified detail of the box in C. Image adapted from Andrews et al., 2019 under CC-... J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R., Yao, L., Yaniv...protocol is a good place to get started.  One important consideration when deciding whether you can do...specific considerations to keep in mind. Most importantly, make sure you are using primary antibodies from...
  5. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...please sign up here.        New toolkit for N or C terminal tagging of proteins for degradation using...has created a new toolkit of plasmids for the N- or C- terminal tagging of proteins with mAID and other ...cytidine deaminase  Target-AID-NG, which mediated a C to T conversion in the target sites adjacent to an...Earlier this year, the Hegemann lab and collaborators report their discovery of a new family of seven ChRs derived...
  6. Bacterial Expression Systems

    Type
    Collection
    ...peptides include: Epitope tags: 6xHis, Flag, Strep II, c-Myc, HA, V5, GST Solubility tags: MBP, SUMO, TrxA,...reconstructed) BiFC Lynne Regan 168257 168472 pMRBad-C-wtBlc pET11a-N-DiB2 DiB2 (reconstructed) BiFC Jens...coli Cynthia Collins 112197 pANY3 Temperature (42 °C) Escherichia coli Yingfeng An Additional Addgene Controlled...Optogenetics for E. coli Return to top Reporter Plasmids Reporter plasmids can be used to detect events...proteins and specific DNA sequences. ID Plasmid Reports Reporter Mechanism Expression Species PI 14474 pRU1161...protein interactions, controlled gene expression, reporter plasmids and genome engineering of bacteria. Plasmid...Visualization Protein Interactions Controlled Expression Reporter Plasmids Genome Engineering You may also like…...
  7. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C., & Telesnitsky, A. (2008). Pseudodiploid Genome...A., Hunter, E., & Kappes, J. C. (1997). Functional RT and IN incorporated into HIV-1 particles independently...34372494. Prussin, A. J., Garcia, E. B., & Marr, L. C. (2015). Total Virus and Bacteria Concentrations in...
  8. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...target sites that code for amino acids close to the C’ terminus of the protein, to maximize the chances ... absence of dsDNA breaks (Rees et al., 2018). For C>T and A>G base editors, the intended edit must be ...bystander edits are possible if there is another target C or A in the window. Another technology, prime editing...doi.org/10.1038/s41587-020-0561-9 Dempster JM, Pacini C, Pantel S, Behan FM, Green T, Krill-Burger J, Beaver...JL, De Los Angeles A, Mali P, Aach J, Kim-Kiselak C, Briggs AW, Rios X, Huang P-Y, Daley G, Church G (...sequence are important considerations for designing your gRNAs. For indels, it's not so important what location...nom. Me love gRNAs! Comic by Maya Kostman.   Important considerations before you start an experiment ...
  9. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ... Sellnow, R. C., Boye, S. L., Coberly, B., Bennett, A., Agbandje-McKenna, M., Sortwell, C. E., Hauswirth...Backer, M. W. A., Brans, M. A. D., Luijendijk, M. C., Garner, K. M., & Adan, R. A. H. (2010). Optimization...Dudek, A. M., Pillay, S., Puschnik, A. S., Nagamine, C. M., Cheng, F., Qiu, J., Carette, J. E., & Vandenberghe..., J. Y., Han, J. H., Lee, H., Han, Y. E., Rah, J. C., & Park, H. (2020). Labeling dual presynaptic inputs...injection site  (Kanaan et al., 2017), axonal transport  (Murlidharan et al., 2014), and postsynaptic ... system, remember that some serotypes can be transported retrogradely and/or anterogradely along the axon...et al., 2014; Nieuwenhuis et al., 2021). It is important to note that this expression pattern can be affected...
  10. Antibodies 101: Normalization and Loading Controls for Western Blots

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    Blog Post
    ...independent blots. B–C) Graphs showing the relative intensity of the β-actin (B) or total protein (C) on the membrane...quantification in panel B is not especially linear. In panel C, two different methods of quantifying total protein...linear. Note the different y-axes between panels B and C, indicating that β-actin is less sensitive as well...BioRender.com. Loading controls are even more important for semi-quantitative western blots. No pipetter...in the same lane, and the resulting fraction is reported. Figure 2: Normalization with loading controls...equally strong, so the normalized value simply reports which POI band is stronger. The right gel has exactly...abundance on your membrane, which would be directly reported by band intensity. Researchers use later steps...
  11. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ... was reduced by growing seed stocks at 30°C compared to 42°C. Screen multiple colonies to isolate the ...19763938. Maucksch C, Bohla A, Hoffmann F, Schleef M, Aneja MK, Elfinger M, Hartl D, Rudolph C. Transgene expression...Growing the culture at a lower temperature such as 30°C or optimizing the incubation time can help maintain...long-read sequencing techniques, such as Oxford Nanopore sequencing or PacBio SMRT sequencing, (which differ...long-read sequencing (1-25 kb), such as Oxford Nanopore sequencing or PacBio SMRT sequencing. These longer... plasmid multimerization troubleshooting tips. Portions of this figure were made with BioRender. ...AE. Generic plasmid DNA production platform incorporating low metabolic burden seed-stock and fed-batch...
  12. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...identification of FLPe which has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998). FLPe’s performance... receptor (ER) ligand binding domain (LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2 #14797). These...FLP-FRT. In addition, a variety of means to spatiotemporally control FLP and Cre expression have been developed...These differences in recombinase efficiency are important to keep in mind when deciding whether to use the... only occur at a specific time point? To allow temporal control, ligand regulated Cre and FLPe recombinase...control of Cre recombinase. After in-utero electroporation, this plasmid mix results in sparse labeling...
  13. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    Blog Post
    ...the reprogramming factors- OCT3/4, SOX2, KLF4, and C-MYC [1]. For the derivation of clinical grade hiPSCs...vectors that encoded six factors - OCT3/4, SOX2, KLF4, C-MYC, LIN28 and SV40 large T antigen (SV40LT). The ...of TP53 enhances iPSC generation as does replacing C-MYC with L-MYC [10, 11]. While the mechanisms underlying...understood, the addition of an shRNA for P53 and replacing C-MYC with L-MYC in the episomal reprogramming cocktail...into cells by electroporation. hiPSC colonies are obtained by 10 days after electroporation, most of which...over a decade back when Yamanaka and colleagues reported that it is possible to reprogram differentiated...generation of human iPSCs (hiPSCs) now provides an opportunity to develop and use patient-specific somatic cells...
  14. Trimmer Lab NeuroMab Collection

    Type
    Collection
    ...IgG2a 177559 Anti-c-Fos [N486/32R] c-Fos Human Mouse IgG2a 177560 Anti-c-Fos [N486/76R] c-Fos Human Mouse...IgG2a 177520 Anti-LRRK2/Dardarin, C-terminus [N241A/34R] LRRK2/Dardarin, C-terminus Human Mouse IgG2a 177521...glutamate receptor Mouse Mouse IgG2a 206619 Anti-c-Fos [N486/25R] c-Fos Human Mouse IgG2a 206620 Anti-Kv1.4 K...220411 LRRK2/Dardarin kinase, C-terminus [N241A/58R] LRRK2/Dardarin kinase, C-terminus Human Mouse IgG2a ...VAPA [N479/111R] VAPA Rat Mouse IgG2a 220414 c-Fos [N486/93R] c-Fos Human Mouse IgG2a 220415 Cavalpha2delta2...] N483/126 Rufy3 Mouse Mouse 182090 C-Fos scFv [N486/32] N486/32 C-Fos Human Mouse 182091 Mortalin/GRP75...84 scFv Rufy3 Mouse Mouse 190560 c-Fos scFv [N486/76] N486/76 scFv c-Fos Human Mouse 190561 BKbeta2 K+...
  15. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    ...21356876. 5. J. Chow, Z. Marka, I. Bartos, S. Marka, J. C. Kagan, Environmental Stress Causes Lethal Neuro-Trauma...PubMed PMID: 28704652. 6. V. Rodríguez, D. Didiano, C. Desplan, Power tools for gene expression and clonal...Neurosci. 9, 703–709 (2006). PubMed PMID: 16582903. 8. C. J. Potter, B. Tasic, E. V Russler, L. Liang, L. Luo...you to systematically study gene function with temporal control and cell-type specificity! The Gal4/...
  16. Antibodies 101: Flow Cytometry Controls

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    Blog Post
    ...starting point for compensation (Fig. 1B-C). However, it’s important to note that because you’re using beads... single color control for the APC channel. B) and C) A flow plot showing an APC single color control prior...prior to compensation (B) and after compensation (C) with APC-Cy7.   FMOs FMO stands for fluorescence...
  17. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    ...and Rose C. St Pierre, founders of STEAMid. Internships open doors to a web of opportunities that the ...guest bloggers Roodolph (Roo) P. St Pierre and Rose C. St Pierre! Roodolph (Roo) P. St Pierre is currently...populations. Roo is the co-founder of STEAMid.       Rose C. St Pierre is currently a graduate student at Northeastern... not deter any student from applying to the opportunities of their dream.  To help students focus on the...below:  1. Eligibility check Scrutinize every opportunity prior to starting an application to assess your...unless you are applying for a Federal Government opportunity, which tends to be more detailed. Beyond the ...draft a personal statement, which is the perfect opportunity to paint a comprehensive picture of your academic...
  18. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    Blog Post
    ...to fuse the activators instead of at the protein’s C terminus. With that new design in place, Konermann...Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki...Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann ...nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop...knockout screening). As the Zhang lab's Nature paper reports, “Here we have shown that the SAM system is robust...size of the backbone, transformations require electroporation. References & Further Reading: For more ...
  19. Deep Dive: qPCR

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    Blog Post
    ...Annealing temperature should be 60-65°C For qPCR, the important primer variable is the annealing temperature... the primers.  Melting temperature should be ~60°C (between 50 and 65, with no more than 3 degrees between...curve will show multiple peaks.    (a) (b)  (c)   Fig. 1: Melt curves showsing (a) a single peak...dimers. (b) A shoulder peak indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits...02217-xTaylor, SC; Nadeau, K; Abbasi, M; Lachance, C; Nguyen, M; Fenrich, J. The Ultimate qPCR Experiment...Zuo J, Dong X, Zhang M, Hu G, Fang F. Fudenine, a C-terminal truncated rat homologue of mouse prominin...step. Scorpion chemistry in particular has been reported to work well in the Lightcycler while being relatively...
  20. CRISPR 101: Non-Homologous End Joining

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    Blog Post
    ...Oh S, Tomkinson AE, Hendrickson EA, Giovannangeli C, Jasin M, Brunet E (2014) Chromosomal Translocations.../10.1016/j.molcel.2014.08.002 Ran FA, Hsu PD, Lin C-Y, Gootenberg JS, Konermann S, Trevino AE, Scott DA... Yan M-Y, Li S-S, Ding X-Y, Guo X-P, Jin Q, Sun Y-C (2020) A CRISPR-Assisted Nonhomologous End-Joining...process involve repairing chromosomal DSBs. It is important to know that cells possess two major repair pathways...chance of causing a frameshift mutation. Of some importance, the deletion can be less heterogeneous when ...
Showing: 101 - 120 of 400 results