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We narrowed to 65 results for: c myc del 2

Showing: 41 - 60 of 65 results

  1. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...repository of tools for SAR-CoV-2 research. Two new antibodies, Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17...Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86], target distinct epitopes on the SARS-CoV-2 nucelocapsid ...Anti-SARS-CoV-2 Antibodies here! Terry, J.S., et al. (2021). Development of a SARS-CoV-2 nucleocapsid ...prime editor PE7 Antibodies to facilitate SARS-CoV-2 research New viral vector preps of Voltron2 voltage... incorporates La’s N-terminal domain, PE7 (Figure 2).  PE7 shows enhanced editing efficiencies in a number...editor. Find PE7 plasmids here!     Figure 2: PE7 improves prime editing. A) Schematic of PE7 prime...PMID: 38570691. Antibodies to facilitate SARS-CoV-2 research   By Ashley Waldron Ready-to-use recombinant...

  2. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ... of components, such as your CRISPR enzyme or Pol 2 promoter. The options available in each class, or ...Cas proteins, guide cassettes, promoters, N’ and/or C’ terminus, and selection markers. Need fewer components...overhang specific to the type of module it is (Pol 2 protomer, Cas protein, ect...). Whatever individual... (KO, CRISPRa, CRISPRi, editing)   Figure 2: Selecting module fragments on the Fragmid website...Griffith, A. L., Szegletes, Z. M., Wen, B., Kraus, C., Miller, N. W., Steger, R. J., Escude Velasco, B....depicting available destination vectors for various delivery methods, along with compatible inserts. All destination...because of their broad applicability in different model systems. What’s the error rate? The Doench lab tested...

  3. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ... Streptococcus , Streptomyces , and others) Drosophila Plants (monocots and dicots) C. elegans Yeast (...transcribed to make the pre-CRISPR RNA (pre-crRNA). (2) The pre-crRNA is processed into individual crRNAs...2015. Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain effector) Type I (Cas3) Type ... with unprecedented speed and specificity. Figure 2: An overview of CRISPR and NHEJ/HDR. The Cas9/gRNA...Zebrafish Xenopus References Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero...23287718 Dalvai M, Loehr J, Jacquet K, Huard CC, Roques C, Herst P, Côté J, Doyon Y. 2015. A Scalable Genome-Editing-Based...Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann ...

  4. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...arrays (e.g. for spacers A, B, and C, you could obtain arrays A, B, C, AB, AC, BC, and ABC). Nielsen Lab...express 2-4 gRNAs, with four being the ideal number. First you generate four unique kanamycin-resistant...all but one of your gRNAs into spectinomycin-resistant plasmids pX330S-2 to pX330S-(last gRNA number). ...illustrated in the schematic below.   Figure 2: gRNA target sequences (colored rectangles) are cloned...the total number of gRNAs you wish to clone, from 2-7. For example, if you’re expressing 4 gRNAs, you’...Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through Golden Gate or Gibson Assembly. gRNAs...Multiplex Plasmids: These plasmids allow expression of 2-5 gRNAs in zebrafish. Custom destination vectors are...

  5. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ..., 146(2), 318–331. https://doi.org/10.1016/j.cell.2011.06.019  Wang, H., Yang, H., Shivalila, C. S., ...extraction solution, runs the protocol, then uses around 2–4 µl in each 50 µl PCR reaction. The gDNA extracted...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.2013.04.025  Yang, H., Wang, H., Shivalila, C. S., Cheng, A. W., Shi, L., & Jaenisch, R. (2013)....marker such as puromycin, you can (also) select for the positive colonies by puromycin selection. You ...for each locus into PX330, then co-transfect or delivery the two plasmids together into your cells. If ...

  6. mTOR Pathway

    Type
    Collection
    ...PKC beta PKC delta PKC epsilon PKC gamma PKC eta PKC iota PKC theta PKC zeta Protein kinase C Protor Also... Apr 13;149(2):274-93. doi: 10.1016/j.cell.2012.03.017. PubMed PMID: 22500797 . Rapamycin: one drug, many...known as RPS6KB1; Ribosomal protein S6 kinase B1 TSC1/2 TSC1 TSC2 Tuberous sclerosis VHL Von Hippel-Lindau...Rictor RPTOR independent companion of MTOR, complex 2 SGK Serum/glucocorticoid regulated kinase 1 Return...Resources The m echanistic or m ammalian t arget o f r apamycin (mTOR) is a key metabolic regulator controlling...threonine kinase 11 mTOR Mechanistic target of rapamycin NF1 Neurofibromin 1 PRAS40 Also known as AKT1S1...associated protein 1 mTOR Mechanistic target of rapamycin p53 TP53; tumor protein p53 PDK1 Pyruvate dehydrogenase...

  7. 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...neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic of the interaction between the sender cell...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...CRISPR Plasmids and Resources page.  Plasmids for delivering CRISPRi components by a single AAV vector are...repression, and editing with a single Cas9-VPR in Saccharomyces cerevisiae. To control Cas9 activity and influence...

  8. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome would restore...positive cells (Martin et al., 2019). Figure 2: Editing efficiencies using base editing reporters...codon that is converted to a ‘TAC’ or ‘TAT’ after a C-to-T base editing event. This edit changes the histidine... ATG codon from an ACG codon using a base-editor (C>T conversion) they could induce the translation of...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464... of plasmid delivery to a cell. In contrast, TREE provides a read out for plasmid delivery and subsequent... a mScarlet reporter, luciferase reporter, and neomycin resistance reporter. Image adapted from Katti ...

  9. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...research experiences to undergraduate students (1, 2), but educators face significant challenges in designing...procedure adds both V5 and His6 epitope tags to the C-termini of Met proteins, which can be used to confirm... by western blotting in other experiments (Figure 2).   Complementation reveals conservation of function...Biology Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology...abnormalities due to the deletion (i.e. the plasmid complements the deletion). If the phenotype is corrected...budding geneticists: A primer on the Saccharomyces cerevisiae model system. Genetics 197, 33-48 (2014)....young geneticists: A primer on the Schizosaccharomyces pombe model system. Genetics 201, 402-423. (2015...

  10. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...Currently available from the Kim lab are 2 presynaptic and 2 postsynaptic targeting mGRASP plasmids. Additionally...the mitochondrial targeting sequence of cytochrome c oxidase subunit VIII, so that they can be imported... cells were pulsed with 2µg/ml of Doxycycline for 2 hours, and imaged on the Keyence BZ-9000 fluorescence...work! Robinson et al., Dev Cell 2010 Feb 16;18(2):324-31. Cheeseman et al., J Cell Sci 2013 May 1...promotes the formation of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated... of three sets of cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly...1 modules can be similarly assembled into a Level 2 vector, thus forming a functional genetic circuit....

  11. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...expressing cells exhibited a 2-4 fold increase in the number of endogenous indel edits when compared to unsorted...Denham lab made several other small changes creating 2 other versions but these did not surpass GEmCherry2...fluorescence and in-frame fluorescence.  Figure 2: The GEmCherry reporter. Image from Højland Knudsen...these methods a try.  References Højland Knudsen C, Ásgrímsdóttir ES, Rahimi K, Gill KP, Frandsen S, ...stably integrate a reporter gene that contains a puromycin resistance gene followed by the target site (the...genome edit:  A donor plasmid that contains a puromycin resistance gene linked to an in-frame H2B-GFP....which depending on the nucleotides inserted or deleted, can result in an in-frame shift of mCherry and...

  12. Tips for arabidopsis transformation

    Type
    Blog Post
    ... Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed...be used for promoter swapping and generating N and C terminally tagged fusion proteins with fluorescent...PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ... plant selection markers (basta, hygromycin, kananamycin, tunicamycin) (Nakagawa et al., 2007). Alternatively...Stanford University. Arabidopsis is a fantastic model organism for many reasons, not the least of which...process right the first time. I’ll lay out some guidelines here. A detailed transformation protocol, as ...

  13. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression  m15 pREP4*, ** Kanamycin (pREP4)...conditions. Many recombinant proteins express better at 30°C or room-temperature, which is accomplished by growing...growing your culture to the desired density at 37°C and reducing the temperature or moving it to a bench-top...lon protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce proteolysis...toxic or insoluble proteins Origami2 (DE3)** Streptomycin and Tetracycline Contains highly active thioredoxin...

  14. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...partial gene deletion, overexpression (by promoter substitution), and tagging at either the N- or C-terminus...Pringle’s former lab at UNC Chapel Hill. Bähler et al.(2) describe a modular collection of plasmids for a wide...protein-protein interactions via FRET. Sidae Lee, Wendell Lim, and Kurt Thorn at UCSF have recently developed...C-terminus (3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary set of plasmids for use in...

  15. CRISPR Plasmids - Drosophila

    Type
    Collection
    ...gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources ...plasmid Bullock and Port 49409 pCFD2-dU6:2gRNA dU6:2 BbsI Injection or in vitro transcription Virmilion...frequently causes insertions or deletions (indels) in the DNA. Indels often lead to frameshifts, creating...is capable of installing targeted insertions, deletions, and all possible base-to-base conversions using...plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance Co-expressed Cas9 Depositing lab Cas9...Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin yes, cut Ji-Long Liu 45946 pU6-BbsI-chiRNA dU6...

  16. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...habitats - even 50 meters from Old Faithful in 60 °C soils. Dendrobatidae (Lauren O’Connell)  Poison frogs...Organisms, Part 2 ...our other model organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms...the spiny purple sea urchin. It’s not a standard model organism, but it was easy to harvest eggs and sperm...So, think beyond Drosophila melanogaster or Saccharomyces cerevisiae (both fine research organisms) -  ...

  17. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Recommended materials C.2 Annealing oligos C.3 Digesting pLKO.1 TRC-Cloning Vector C.4 Ligating and transforming...ligation, mix: 2 μL annealed oligo from step C.2 20 ng digested pLKO.1 TRC-cloning vector from step C.3 (If you...at 37°C, 5% CO 2 overnight. Day 2: b. The target cells should be approximately 80-90% confluent. c. Dilute... your gene B.2 Order oligos compatible with pLKO.1 C. Cloning shRNA oligos into pLKO.1 C.1 Recommended...VWR: #7177-48-2. Use at 100 μg/mL. Carbenicillin VWR: #80030-956. Use at 100 μg/mL. C.2 Annealing Oligos...Reverse oligo 5 μL 10x NEB buffer 2 35 μL ddH 2 O Incubate for 4 minutes at 95°C in a PCR machine or in a beaker...buffer 1 1 μL AgeI add ddH 2 O to bring to 50 μL final volume Incubate at 37°C for 2 hours. Purify with Qiaquick...

  18. Molecular Biology Reference

    Type
    Guide
    ... T H A, C, or T K G or T M A or C N A, T, C, or G R A or G S C or G V A, C, or G W A or T Y C or T Amino...dissolved in dH 2 0. Addgene recommends making 1000X stock solutions and storing aliquots at -20°C. To use, ...Thymine (T), Cytosine (C) and Guanine (G). In the double helix A always pairs with T and C always pairs with...Adenine C Cytosine G Guanine T Thymine U Uracil Single Letter Code: Ambiguous bases Nucleobase B C, G, or...Acid Sequence FLAG DYKDDDDK HA YPYDVPDYA His HHHHHH Myc EQKLISEEDL V5 GKPIPNPLLGLDST Xpress DLDDDDK or DLYDDDDK...thymine, cytosine, and guanine (abbreviated to A, T, C, and G respectively) that are organized into a double...complementary, strand. Specifically, A pairs with T and C pairs with G. During replication, DNA unwinds and ...

  19. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ... Multiple Introns.” Plant Communications, vol. 2, no. 2, Elsevier BV, Mar. 2021, p. 100135. Crossref, ...Constructs.” PLoS ONE, edited by Jean Peccoud, vol. 6, no. 2, Public Library of Science (PLoS), 18 Feb. 2011, p... vol. 5, no. 10, American Chemical Society (ACS), 2 May 2016, pp. 1059–69. Crossref, https://doi.org/10.1021...doi.org/10.1371/journal.pone.0197185. Chamness, James C., et al. “An Extensible Vector Toolkit and Parts Library...of Plant Genomes.” The Plant Genome, vol. 16, no. 2, Wiley, 9 Mar. 2023. Crossref, https://doi.org/10.1002...Crossref, https://doi.org/10.1186/s12870-020-02388-2. Stuttmann, Johannes, et al. “Highly Efficient Multiplex...Crossref, https://doi.org/10.1038/s41598-019-46171-2. Resources on Addgene.org Modular Cloning (MoClo...

  20. CRISPR References and Information

    Type
    Collection
    ...coli, B. subtilis ), yeast ( S. cerevisiae ), worm ( C. elegans ), fruit fly, zebrafish, mouse, rat, and ...Link opens in a new window) An encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9-Blast and lentiGuide-Puro...off-target binding. Currently supports over 20 model and non-model invertebrate species. Developed by the Kate... of CRISPR plasmid kits. CRISPR Software Sanger Indel Analysis ICE (Inference of CRISPR Edits) This open...al. (2019) (Link opens in a new window) . MAGeCK Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout...a new window) A webtool that uses deep learning models for SpCas9 gRNA design and efficiency prediction...
Showing: 41 - 60 of 65 results