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We narrowed to 45 results for: c myc del 2

Showing: 21 - 40 of 45 results

  1. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...combined the system with Flp/FRT, as reviewed by del Valle Rodriguez et al.   LexA/lexAop A second binary...Find plasmids from this publication at Addgene. del Valle Rodriguez, Alberto, Dominic Didiano & Claude...engineering applications in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197-214. PubMed PMID...also shown that the Q system is also functional in C. elegans. Subsequent work by the Luo and Potter labs...immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425...23995928; PMCID: PMC3811519.Seo SO, Schmidt-Dannert C. Development of a synthetic cumate-inducible gene ...Targeted Gene Expression in Drosophila.” Genetics 186(2) (2010): 735-55. PubMed PMID: 20697123. PubMed Central...

  2. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ... 79628 (C) Cv (capsid) N and C 79633 (N) and 79641 (C) NS1 (var: W98G) N and C 79635 NS3 C 79637...79637 NS2B C 79636 NS4A N 79640 NS4B C 79639 NS5 C 79632 (N) and 79631 (C) PrM N and C   Detecting...tagged BioID2 for N-terminal fusions and Myc tagged BioID2 for C-terminal fusions. Kim, et al. Mol Biol...expressed gRNAs respectively) and 2) the size of the deletion. Smaller deletions generally occurred at higher...backbone, which expresses the gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene...backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used GMAP assembly...engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand...

  3. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...difficult to grow. Growth Temp Choose from 30 °C, 37 °C, or room temperature. Growth Strain Please...our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer team...columns are shown in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down...frog), D. rerio (zebrafish), D. melanogaster (fly), C. elegans (nematode), S. cerevisiae (budding yeast)...Filling out and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first... gentamicin, hygromycin, kanamycin, nourseothricin (clonNat), spectinomycin, streptomycin, tetracycline...exceptions to our standard workflow.  Table 1: Guidelines for the data fields in the deposit spreadsheet...

  4. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    Published
    July 29, 2025, 1:15 p.m.
    ...Oligo 2 to screen. A 667-bp product confirms successful insertion. Step 6: Confirmation Pick 2–3 positive...doi.org/10.1038/s41587-019-0337-2  Molla, K. A., Sretenovic, S., Bansal, K. C., & Qi, Y. (2021). Precise ...can use TEEP — a tool based on animal data. Step 2: Prepare Oligos Oligo 1: Add 'tcaa' to the 5′ end...your forward guide sequence (see Figure 1). Oligo 2: Add 'ggcc' to the 5′ end of your reverse complement...Primer 92F (5′-cattacgcaattggacgacaac-3′) and Oligo 2 (specific to your target) to screen colonies via PCR...Maher, M. F., Nasti, R. A., Vollbrecht, M., Starker, C. G., Clark, M. D., & Voytas, D. F. (2019). Plant gene...Gao, Q., Liu, S., Zhang, X., Hu, W., Wang, Y., Han, C., Li, D., & Wang, X. (2025). Transgene- and tissue...

  5. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    Published
    Oct. 10, 2023, 1 p.m.
    ...jcm.23.5.959-961.1986 Lobstein, J., Emrich, C. A., Jeans, C., Faulkner, M., Riggs, P., & Berkmen, M. (2012...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...References Hwang, J. M., Piccinini, T. E., Lammel, C. J., Hadley, W. K., & Brooks, G. F. (1986). Effect...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I ...Slow transformation   Spectinomycin/streptomycin Spectinomycin and streptomycin both inhibit protein synthesis...isn’t a widely popular selection agent. Pros: Stable (spectinomycin). Cons: Cost (spectinomycin) or stability... for transformations. Con: Expensive.   Kanamycin Kanamycin, or kan, is an aminoglycoside which inhibits...

  6. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...Sadowski. 1985. “The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently to...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 and Cre for additional control   Figure 2: Expression of a morphological marker (eYFP) and synaptic...Sternson. 2008. “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range ...PMID: 18614669. PubMed Central PMCID: PMC2593125. 2. Buchholz, F, P O Angrand, and A. Francis Stewart....

  7. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ..., while pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ... of cytidine and adenine base editors that enable C-to-T and A-to-G base substitutions respectively. Substitutions...:723–728 . https://doi.org/10.1021/sb500351f Dong C, Fontana J, Patel A, Carothers JM, Zalatan JG (2018... https://doi.org/10.1016/j.cell.2013.02.022 Ronda C, Pedersen LE, Sommer MOA, Nielsen AT (2016) CRMAGE...Vridhambal GS, Rivera AJ, Montgomery SA, Fortier L-C, Barrangou R, Theriot CM, Ousterout DG (2020) In Vivo...

  8. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...El-Bakkoury, M., Hamacher, T., Cappellaro, C., Vilarino, C., Fleischer, C., Ellerbrok, H., Kamuzinzi, R., Ledent... systems available in model organisms, including bacteria, alternative fungi (C. albicans, pC2HB) (7) ...retain their individual activities when separated (2), and that DBD and TAs from different systems could... prey libraries can be screened using both N- and C-terminal fusions of these proteins. This way both ... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...Two hybrid systems were developed in Saccharomyces cerevisiae in 1989 and are still used extensively ...

  9. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...fluorescent state make them useful for 2-color imaging with green PA-FPs. Table 2: Properties of Selected Reversible... conventional FP imaging. PA-FPs are divided into 2 main categories - those with irreversible and reversible... forms of the chromophore which contribute to the 2 peaks of wtGFP’s excitation spectrum- a major 397 ... / N99K / S148L / I165V / Q167P / L169V / I203R) (2) and PA-mRFP1(S146H / I161V / I197H) (3) were derived...mEosFP’s chromophore requires a temperature below 30°C and this is not ideal for experiments in mammalian...can form it’s chromophore at a more comfortable 37°C (5). Dendra2 photoconverts from green to red fluorescence...with two distinct wavelengths of light (see Table 2). The best known reversible PA-FP is the naturally...

  10. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ...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...

  11. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ... 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...

  12. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...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...

  13. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ..., 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 ...

  14. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    May 22, 2018, 1:38 p.m.
    ...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...

  17. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...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....

  18. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...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...

  19. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ... 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 ...

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

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...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...
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