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We narrowed to 337 results for: c rel

Showing: 141 - 160 of 337 results
  1. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...Zhi Ying Chen, Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle... generating iPSCs. The generation of iPSCs is relatively simple in concept: ectopically express a cocktail.... The advantages of this method are that it’s relatively simple to implement and doesn’t require time-...a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous MicroRNAs." Plos One 11, no...Overall, integrating viral vectors efficiently and reliably generate iPSCs, but they have several safety concerns...important control. Additionally, Sendai virus doesn’t rely on promoters for transcriptional regulation, so ...Check Out Our Interview with Stem Cell Researcher Darrell Kotton Four Factors that Differentiate the Stem...
  2. 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... colocalization factor to assess the spatial relationship between the targets. Or, it might be sufficient...
  3. COVID-19 Resources

    Type
    Collection
    ...Andersen, K. G., Rambaut, A., Lipkin, W. I., Holmes, E. C., & Garry, R. F. (2020). The proximal origin of SARS-CoV..., Wu, N. H., Nitsche, A., Müller, M. A., Drosten, C., & Pöhlmann, S. (2020). SARS-CoV-2 Cell Entry Depends...the knowledge and understanding of COVID-19 and related coronaviruses. Addgene assisted this effort by ...tools that are useful for COVID-19 related research. Additionally, we have linked to collections...articles, protocols, and other resource collections related to COVID-19 that may be of use to scientists. Antibodies...entry of SARS-CoV and SARS-CoV-2. Find plasmids related to these genes below. ID Plasmid Description Industry... in the body. For a full list of cytokines and related plasmids see our Immunology Resource Guide . Return...
  4. Bacterial Expression Systems

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    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...interactions using fluorescence microscopy. FRET relies on the principle that when a donor fluorophore ...for a selection of mammalian and bacterial FRET-related vectors and standards. The Hoschschild Bacterial... that vary in strength and that can be used to reliably drive expression of a gene of interest in E. coli...
  5. 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... and how these pathways work, as this could be relevant when planning your experiment. Here, we’ll discuss...repair is that the Cas9 protein doesn’t immediately release from the broken end after cleavage, which may interfere...
  6. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    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 ... not managed to create a system to allow us to reliably activate essentially any gene,” she says. The ...tools because obviously, unlike the mouse, we can’t rely on genetic engineering to optimize humans for CRISPRi...
  7. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...should the double strand break (DSB) be positioned relative to where the homology ends? A9: The Zhang lab ...10.1126/science.1231143  Elliott, B., Richardson, C., Winderbaum, J., Nickoloff, J. A., & Jasin, M. (1998....1128/mcb.18.1.93  Ran, F. A., Hsu, P. D., Lin, C., Gootenberg, J. S., Konermann, S., Trevino, A. E....j.cell.2011.06.019  Wang, H., Yang, H., Shivalila, C. S., Dawlaty, M. M., Cheng, A. W., Zhang, F., & Jaenisch...j.cell.2013.04.025  Yang, H., Wang, H., Shivalila, C. S., Cheng, A. W., Shi, L., & Jaenisch, R. (2013)....
  8. Click Biology: How to Click and Play to Explore Biology

    Type
    Blog Post
    ...Cysteine-less, fast split intein; N-terminal CLm and C-terminal Aes gp41-1 Split inteins A small, fast ...attB2r-SNAP-attB3 SNAP-tag Gateway entry clone to add C-terminal SNAP-tag Promega Plasmid Collection HaloTag...PCV-Cas9, Cas9-PCV HUH-tag Cas9 plasmids with N- or C-terminal PCV2 HUH-tag; increase homology-directed ...Click Biology describes the search for robust, reliable reactions, using only nature’s building blocks... are many convenient tools out there, not just relating to proteins but also covalent reactions to DNA...Click biology highlights the opportunities from reliable biological reactions. Nature Chemical Biology....
  9. Plasmids 101: Dimers and Multimers

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    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...overlap with the band corresponding to the nicked or relaxed conformation of the plasmid monomer (see this article...
  10. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    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...containing FLEx technology. Read the genOway press release for more information. References 1. Atasoy, D.,...
  11. PCR Applications for Copy Number Variation Assays

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    Blog Post
    ... one-copy ‘housekeeping’ gene), and quantified relative to a 'normal' sample with known copy number set...nice thing about qPCR is that since it measures relative quantification - that is, since you’re looking...looking at the amount of DNA in your sample relative to your normal control - you can set up your calculations...provides absolute quantitative data, instead of relative quantification.  Even though no one option is ... M., Sim, B., Mustafa, M., Nik Yusof, N. K., Lee, C. K., Abu Bakar, S., AbuBakar, S., Hollox, E. J., &...
  12. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...at 95 °C, a common lysis temperature, so consider a longer incubation (10–20 minutes) at 70 °C, or a truly... Additionally, protein lysis should be done at 4 °C or on ice, whichever is more practical. If your lysing... truly lengthy incubation (30–60 minutes) at 37 °C, if you suspect aggregation in your prep. If your proteins...protein-based blockers degrade fairly quickly, even at 4 °C, so when in doubt, make up a fresh batch.  Antibodies...and time. Though the most common conditions are 4 °C overnight or room temperature for 1–2 hours, there...sample loaded Incubate primary antibody at 4 °C overnight Incomplete washing Ensuring wash is... may need to sonicate your lysates in order to release the proteins from their binding. If you’re seeing...
  13. RNA Extraction Without A Kit

    Type
    Blog Post
    ...store them in a -80°C freezer to prevent RNA degradation. They will degrade in a -20°C freezer.   RNA extraction...DEPC. Alternatively, glassware can be baked at 180°C for at least 4 hours.   If your final RNA sample(s...As with DNA isolation, scientists commonly rely on RNA isolation kits to make their life easier. Recently...resources on the Addgene blog Read blog posts related to plasmid protocols Find protocols for purifying...
  14. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...cloning method with high throughput capability. Engler C, Kandzia R, Marillonnet S. PLoS One 2008;3(11):e3647..., Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic...2011 Apr 14. PubMed.   Golden Gate cloning. Engler C, Marillonnet S. Methods Mol Biol. 2014;1116:119-31...cloning efforts. Golden Gate cloning technology relies on Type IIS restriction enzymes, first discovered... destination vector + insert may spontaneously religate, this transient construct retains functional Type...
  15. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...interactions New auxin inducible degron system for C. elegans New CRISPR plasmids New items from the viral...tool-kit for the auxin inducible degron (AID) system in C. elegans By Alyssa Cecchetelli The auxin inducible... degron (AID) system has been extensively used in C. elegans since 2015 to conditionally degrade proteins... from Ashley et al., 2021.   The AID system relies on a transport inhibitor response 1 (TIR1) expressed...induce DSBs within minutes. New CRISPR plasmids for Borrelia burgdorferi from Christine Jacobs-Wagner’s lab...
  16. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    ...guest bloggers Roodolph (Roo) P. St Pierre and Rose C. St Pierre, founders of STEAMid. Internships open ...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...certain to choose your recommenders based on your relationships to the person as well as their current sphere...
  17. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ...and T7E1 have been widely adopted due to their relative simplicity and low cost. The problem with these... detection rates mean the investigator can be relatively certain that their samples don’t have off-target...PubMed Central PMCID: PMC4349094. 3. Bell, Charles C., et al. "A high-throughput screening strategy for... 22963480. PubMed Central. 5. Tsiatis, Athanasios C., et al. "Comparison of Sanger sequencing, pyrosequencing...genotyping, is critical to demonstrating causal relationships between genotype and assayed phenotype. Verifying... for validating edits.  Mismatch cleavage assays rely on pairing between the edited strand and wild-type...limit of detection of ~5%. This means they do not reliably detect editing events that occur in less that ...
  18. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

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    Blog Post
    ... promoter, gRNA and pol III terminator. These relatively large cassettes (considering the gRNA itself ... vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from these ...specific genome editing. Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin MJ, Aryee MJ...
  19. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...Williams, Eric J. Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado,... first discovered and characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a...gene expression and functions of key regeneration-related genes in four inexpensive and commercially available...
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