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We narrowed to 457 results for: nsf

Showing: 141 - 160 of 457 results

  1. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...expression vector that is then transformed into a bacterial cell. Transformed cells propagate, are induced... strategy for protein expression consists of transfecting cells with your DNA template of choice and allowing...SV40, CMV, and RSV) for robust expression post-transfection. Mammalian systems can express proteins both... Both methods produce high protein yields if transfection is successful. Some mammalian systems also ...

  2. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...used by respondents (transfection method, amount of each component per transfection etc.). Take Pooran'...an ssDNA guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression...delivery methods. Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA...researchers will have to carry out repeated transfections with the ssDNA guide, although many may find...

  3. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...determined by the different hosts that a plasmid can transfer to, replicate in, and persist in. Some plasmids...easily cloned in different bacteria, and can be transferred to various experimental hosts (Jain and Srivastava...scientific community, as they allow for simplified transformation processes, and can be used by scientists across...conjugative and carries the genes necessary for transfer initiation between bacterial hosts. Also, self-transmissible...Plasmids 101: What is a Plasmid?  Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Tranfection...

  4. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...more stable than the methyltransferase. When a plasmid is lost, the methyltransferase is degraded and/or...ccdB gene die off and a higher percentage of transformants contain the ccdB-less construct. Other toxin-antitoxin...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at ...specific nucleotide sequences. To counter this, methyltransferases add methyl groups to bases at specific nucleotide...

  5. Retrograde AAV viral preps

    Type
    Collection
    ...titering by SYBR green qPCR , confirmation of the transfer plasmid with PCR, purity assessment by silver ...

  6. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...size, multimers may exhibit reduced transformation and transfection efficiency compared to the monomeric...plasmid multimerization is to avoid growing the transformed bacterial culture into the late stationary phase...the linearized band, ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form... Hartl D, Rudolph C. Transgene expression of transfected supercoiled plasmid DNA concatemers in mammalian...

  7. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    ...proteins needed for the virus to replicate,  and the transfer plasmid which contains the viral genome. In today...Replication initiates from the AAV genome ITRs on the transfer plasmid, which contain regions of dsDNA. To generate...out this blog post to learn more about the AAV transfer plasmid! References: Naso MF, Tomkowicz B, ...was engineered Learn about the parts of the AAV transfer plasmid  Resources on Addgene.org Learn about...

  8. What the HEK?

    Type
    Blog Post
    ...1970s by Alex van der Eb in the Netherlands. Transformation of HEK cells with adenovirus 5 DNA was postdoc...replication. To generate a virus, you need only transfect the ‘helper’ plasmids (viral packaging machinery...Protein production  HEKs are extremely easy to transfect in bulk and produce recombinant proteins at high...Resources References Shaw G., et al., Preferential transformation of human neuronal cells by human adenoviruses...

  9. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    ...understand genetic cytoplasmic factors that could be transferred between cells. At the time, these extranuclear...-factors. Like F-factors, R-factors could be transferred between bacteria via cell-to-cell contact; however...pSC102, with EcoRI and selected for E. coli transformants that were resistant to both. When this proved...Chizuko Ogata, and Sachiko Sato. “EPISOME-MEDIATED TRANSFER OF DRUG RESISTANCE IN ENTEROBACTERIACEAE VIII:...

  10. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ...covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR template. Some of the applications...target region. Assembled plasmids can be directly transformed into E.coli for plating, screening and generation..., generating expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time...inserted into a linearized cassette, ready for transformation into yeast that same day.  The field of synthetic...

  11. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...December 2015 we held our first intro to bacterial transformation course (read our blog post on it and to see...consisting of over twenty members and volunteers. Transforming citizens into scientists Anyone can become a...dedicated time at BosLab, he became our bacterial transformation guru and instructor for our intro to biohacking...Addgene and are eager to do our first yeast transformations! Dennis Oleksyuk, Raphael Laurenceau, and Tami...

  12. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    ...Wondering how you can develop more transferable skills? See Developing Transferable Skills During Science Training...advantage over other candidates. Write about your transferable skills Some of the skills you developed might...have presented in some public lectures? These transferable skills could be meaningful for your potential...

  13. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...due to the ease of transduction in difficult-to-transfect cell types ranging from primary to embryonic stem...construct is also reliably higher than traditional transfection and random genomic integration of a plasmid.... is also generally less toxic to cells than transfections, preserving cell viability. Library delivery... with viral vectors instead of a traditional transfection method. This is because viral systems have a...

  14. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...the specific reaction you are running. Transformation Transform a few microliters of your ligation reaction...The PCR could fail to produce a product, the transformation may not result in any cells, or all the colonies...fragments If you don’t get any positive clone, try transforming the ligation mix in different bacterial strains...

  15. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ...hippocampal cultured neurons, imaged 14 days post-transfection, excited at 505 nm (right). Scalebar represents...hydropersulfide-propagating enzymes 3-mercaptopyruvate sulfurtransferase (MPST) which catalyzes the desulfuration of...an enzyme-bound hydropersulfide that is then transferred to thiol-containing acceptors. These hydropersulfides...redox modification, the MPST hydrosulfide can be transferred to the roGFP2 cysteine thiols and change its ...

  16. Working with Nuclear Receptors

    Type
    Blog Post
    ...such as p300 and CBP histone acetyltransferases or CARM1 methyltransferase. This attendant signaling feature...laboratories. The development of the transient transfection platform for assaying transcriptional activity...over the years evolved from chloramphenicol acetyltransferase to luciferase, the basic assay remains widely...

  17. Hot Plasmids February 2024

    Type
    Blog Post
    ...StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific AAV delivery...electroporation or poor cellular entry with other transfection methods. The Junwei Shi Lab addressed this problem... viral vectors for functional intravenous gene transfer throughout the non-human primate brain. Nature...the Gradinaru lab, AAV1-X1, was developed by transferring the 7-mer peptide region from variable region...

  18. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...Cas9 ribonucleoprotein complexes into transformed and non-transformed cells yielded rates of targeted mutagenesis...introduction of DNA into therapeutically relevant, non-transformed mammalian cells. These viral vector systems also...treatment (Stephens et al., 2019). If sufficient gene transfer and knock-in is achieved, producer cells could...

  19. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...over for other applications, like PCR, cloning, transfection, or long-term storage. But what about those ...inverted terminal repeats. We use NEB Stable to transform and prep high quality DNA that contains these ... in a protein expression strain, we suggest retransforming the plasmid into a strain meant for DNA purification...to Quantify Your DNA Learn to Do a Bacterial Transformation Learn How to Perform a Diagnostic Restriction...
Showing: 141 - 160 of 457 results