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Showing: 261 - 280 of 755 results

  1. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ... out a non-essential gene of interest. However, if your gene of interest is essential, a true knockout... suitable PAM sequences nearby or your gene of interest is difficult to target specifically with a gRNA...that contains sequence homology to the locus of interest. To give you an idea of the process, we’ll walk...resistance gene and the targeted exon in the gene of interest (Figure 4). When recombination occurs, the gene...

  2. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ...work anywhere in the genome. Aneesh's research interests include proteomics, machine learning, and visualization...intuition for how genome math helps us to locate interesting addresses in the genome. Let’s now examine how...dies. Order Lentivirus from Addgene If you’re interested in designing your own CRISPR screens for enhancers...alzheimers-affected neurons, or [your cell line of interest], is the foundation of precision medicine. We ...is a collaborative database. Aneesh's research interests span machine learning, proteomics, and user interfaces...

  3. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...of the gene of interest, or expressed from the same transcript as the gene of interest, but after an internal...Placing GFP under the control of a promoter of interest can be used to effectively monitor gene expression... portions of EGFP are fused to the proteins of interest, and when they come into close proximity, the ...be used, when put under control of promoters of interest, to visualize the developmental stage at which...

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

    Type
    Blog Post
    ...pCRISPR: carries a spacer targeting the gene of interest and kanamycin resistance E. coli carrying the...template. Through recombineering, the locus of interest is modified to match the repair template, and ...sequence persists, Cas9 will cleave the gene of interest, inducing a lethal DSB.     This system is distinct... a repair template, using Gibson Assembly or restriction enzyme cloning. Streptomyces bacteria are more...bacteria including in pooled, genome-wide, formats. Interested in knowing more? Check out our Mobile CRISPRi...contain BsaI sites for easy cloning of a gRNA of interest. Cytidine and adenine base editing in Streptomyces...

  5. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...plasmid does not have many restriction enzyme sites. Hosts often have a restriction barrier as a defense mechanism...use of their restriction enzymes (Meyer et al. 1977). So if a plasmid has few restriction sites, it may...R. (1977). Physical and genetic studies with restriction endonucleases on the broad host-range plasmid...

  6. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ... both a sample known to express the protein of interest, such as a sample transiently transfected or stably... stably infected with the protein of interest and a sample that does not. Common negative controls include...c-Myc, GFP or V5 can be used to tag the protein of interest on the C- or N-terminus. In this variation, called...against the tag is used to isolate the protein of interest. The wide availability and high specificity of...protein or interfere with protein interactions of interest. Tagged proteins are typically expressed at much...

  7. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...contains the sgRNA targeting the genomic site of interest. The donor template is supplied separately. To...containing another sgRNA targeting another site of interest can be used to make other edits. Once you have... pTargetF with the ccdB gene containing BsaI restriction enzyme sites at its ends to generate pEcgRNA....replaced by the sgRNA targeting the genomic site of interest. We compared the pCas/pTargetF system with the..., we were able to successfully delete genes of interest in BL21(DE3) using the newly generated pEcCas/...

  8. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...fusion protein. Also, in cases where the protein of interest is relatively small, adding GFP may alter diffusion...protein. Effectively, it equips the protein of interest with an additional protein module that can report...proteins in cells, but not other biomolecules of interest e.g. DNA, RNA, and lipids. Still, these molecules...domains that specifically bind the molecule of interest. For instance, pleckstrin homology domains can...GFP into the coding sequence of the protein of interest. One of the reasons that this works well is that...

  9. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...selected the optimal promoter for your gene of interest, painstakingly cloned into the perfect empty backbone...DNA, important when trying to cut with certain restriction enzymes (ex: ClaI or XbaI) dcm DNA cytosine...DNA, important when trying to cut with certain restriction enzymes that are methylation sensitive. dnaJ...plasmid maintenance, blue/white screening, partly restriction-deficient; good strain for cloning repetitive...methylated, allows for methylation sensitive restriction enzymes to cut the plasmid after preparation....

  10. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...cloning sites for easy cloning of your genes of interest. In order to improve the study of specific neuronal...new AAV approach using regulatory elements that restrict expression to this cell type. To this end, they... ATP1A1 in addition to a researcher’s locus of interest. NHEJ- or HDR-induced mutations in ATP1A1 can ...locus increases the probability that the locus of interest has also been edited! Thus, ouabain selection ...the size of DNA fragments. Depending on which restriction enzyme is used to cut the plasmids encoding the...are shared through Addgene without licensing restrictions. Use these instructions to prepare the PSU ...

  11. Finding Your Perfect Job After University

    Type
    Blog Post
    ...pediatric kidney cancer. This was particularly interesting as I had just completed my thesis on the study... taking on a similar position in your field of interest before you dive straight into a full-time position...for bacterial contamination. It was a really interesting job at first, everything was very new, and everything...to help cover the costs. My MSc courses were interesting and luckily genetics is a sector which has been...the company as well and use my other skills and interests. I was so glad when my future supervisors were...

  12. DIY DNA Ladders from Penn State University

    Type
    Blog Post
    ...developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate...significantly cheaper than commercial ladders. Are you interested in making your own DNA ladder? You can find the...available from Addgene. Learn How to Analyze a Restriction Digest References 1. Henrici, R. C., Pecen, ... Miniprep Protocol Learn How to Analyze Your Restriction Digests Make Your Agarose Gel Other Resources...

  13. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    ...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...age live in a state with at least two abortion restrictions that conflict with scientific evidence. We need...Campus.  He studies viruses including HIV and is interested in global health policy.  He is also chair of...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...science and positively reinforce their scientific interests. Our marches across the world have had an impact...

  14. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...parts in a specific orientation using Type IIS restriction enzymes (Weber et al., 2011). It can be performed...units (TUs) used to express your favorite gene-of-interest, all the way to the complex multi-TU constructs...and tags makes EcoFlex a great option for those interested in recombinant protein purification and metabolic...looking to express high levels of their gene of interest in yeast should also take a look at the Yeast ...stacking, increasing production of a protein of interest, and improving the nutrient profiles within given...

  15. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ...wanted to measure expression of your protein of interest in a single cell? Or perhaps, you need to analyze...fluidics system they pass through a flow cell which restricts the size of the stream and forces the cells to...? However, fusion proteins tag your protein of interest with a fluorescent reporter on the N- or C-terminus... around the specific population that they are interested in and further analyze that subset of cells. .... An immunologist, for example, might be interested in the lymphocytes and “gate” that population for ...

  16. Stepping into a Virtual Internship: My Experience at Addgene

    Type
    Blog Post
    ...always been interested in the ways that I could change the world around me. My strong interest in the intersections... marketing internship at Addgene. Although my interests are very clear, I had no idea how to translate...would be a perfect way of exploring my different interests but also a way of challenging what I am used to...

  17. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    ...multi-color imaging experiments. Interested in epitope tags? Others may be interested in attaching epitope tags... fluorescent proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed...tags to their genes of interest, allowing for easy capture and detection of proteins and complexes, without...

  18. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...the first gRNA using restriction enzyme BbsI and the second gRNA using restriction enzyme BsaI. If you’... of gRNAs Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition...These expression vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs...

  19. An Addgene Summer Internship

    Type
    Blog Post
    ...heavy restrictions on what I could and couldn’t do as an intern. I expected that these restrictions would...choose what gene to focus on. The gene choice had restrictions. It couldn’t be a Covid gene, a toxin, or an...running on a time crunch and knew that if we were to restart the procedure we would not have enough time. During...

  20. Networking and Inspiring Young Scientists at Freshers Fairs

    Type
    Blog Post
    ... students to network, find groups with common interests, and settle into university life. The fairs are...1996, Cambridge Biosoc’s goal has been to promote interest in the biological sciences amongst the student...meetups. The society hopes to help young scientists interested in this field excel and develop their career ...from, and mentor while pursuing your research interests. Scientific societies like those mentioned above...
Showing: 261 - 280 of 755 results