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Showing: 241 - 260 of 755 results

  1. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    ... such as Stbl3s. ITR loss can be detected by restriction digest using enzymes, like SmaI, which have recognition... such as SaCas9, CRISPR gRNAs, or your gene of interest. Promoter The promoter drives the expression of...organism or tissue type-specific, which can help you restrict the expression of your transgene to a specific...result is a permanently flipped transgene. By restricting the availability of Cre, lox sites lets researchers...successful and specific expression of your gene of interest. References Haery L, Deverman BE, Matho KS, ...

  2. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    ...of knowledge or interest in STEM subjects; we generally refer to them as the “interested public.” Subscribers...to Scientific America would be considered the interested public and it would be safe to assume that they...age range, familiarity with the subject, shared interests, socioeconomic and/or racial/ethnic background...example adults 45-55 yrs old who breed or are interested in breeding dogs who also enjoy listening to ...

  3. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    ...need to add unique restriction sites on the ends) and amplify your sequence of interest with Taq polymerase...cloning) is a DNA cloning method that does not use restriction enzymes or ligase, and requires no post-PCR procedures...depicted as a green cloud) functions both as a restriction endonuclease and as a ligase by cleaving and ... confirm the presence of your insert by PCR, restriction digest, or sequencing. Pro tips Do not add 5...

  4. Hot Plasmids: FRET-Based Biosensors

    Type
    Blog Post
    ...one problem: how to get that into the cells of interest,” Griesbeck said. Because they are chemical compounds... to go. Griesbeck is motivated by a particular interest in monitoring the activity and biochemistry of... Technische Universiteit Eindhoven has similar interest in FRET-based sensor proteins, particularly for... doi: 10.1371/journal.pone.0082009. (PubMed) Interested in More Hot Plasmids from Addgene? Check out...newly available plasmids at Addgene. If you're interested in plasmids for in vivo imaging, check out this...

  5. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...interactions is the pull-down assay, wherein a protein of interest can be captured along with its associated partners...chromatin fragments that are bound by a protein of interest are pulled-down and later coupled to high-throughput... the antibody problem is to tag the protein of interest with a small biochemical “epitope tag” such that...generated, each with a different tagged protein of interest, and a universal all-purpose “tag antibody” can...crRNA (guide RNA sequence targeting the gene of interest) and ssODN (tag-encoding donor DNA) using our ...Regenerative Medicine (CRM), Edinburgh. His research interests include optimizing CRISPR/Cas9 genome-editing ...

  6. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...antibodies could thus be interesting drugs. Known drug targets  Another interesting approach is to design...expression dataset are causal for a phenotype of interest. Selected pathways  I attended a great talk when...to calculate which would be the best gRNAs. Interestingly, the libraries do not overlap much, showing ...identify all the alternative names for your gene of interest (that were not found by Green Listed), and test...Karolinska Institutet, in Sweden. He has a particular interest in using molecular tools to study the immune system...

  7. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ... often struggle to understand how a protein of interest actually behaves. How large is it? What parts ...required is that the proteins/nucleic acids/lipids of interest are fluorophore labeled and that these fluorophores...each other. How do you label your biomolecule of interest with FRET fluorophores? When is absolute labeling... FRET acceptor-binding site on your protein of interest: you just don’t know it yet! The poly-histidine... Addgene, where you can insert your protein of interest, resulting in a C-terminal fusion of a tetracysteine...Brigham and Women’s Hospital in Boston, MA. His interests include developing novel fluorescent tools for...

  8. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ...tool that allow scientists to express a gene of interest in a cell or animal model. AAV plasmids contain...cassette, which contains a promoter, the gene of interest, and a polyA signal. Created with biorender.com... an AAV vector plasmid, they flank the gene of interest. A single ITR consists of palindromic arms (A-...identification of non-dominant ITR sequences.  Restriction digests with the enzymes SmaI and SrfI can also...deletions and point mutations will go undetected by restriction digest methods.    Resources and references...

  9. Finding and Joining Your Dream Lab

    Type
    Blog Post
    ...opportunities arise at various times. If the timing and interests align for both parties, he will invite the prospective... Cepko looks for people who are dedicated and interested in science. “Postdoc hires are simpler because...want to do. The experiments must fit with their interests.” George Church selects new hires initially based...full day, gives a 1 hour talk about his or her interests, and spends one-on-one time with the current members...course, the specific research field should be interesting; however, this should (in most cases) not be ...

  10. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ... to introduce frameshift mutations in genes of interest, turning expression off permanently. Viral vectors...promoter (inducible or constitutive), your gene of interest, and often a selectable marker. This vector can...modification of the viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA...mRNA or a sgRNA targeting an exon in your gene of interest, for example. Some customization is required, ... What viruses can do for you Whether you are interested in basic science or clinical therapy—viruses ...

  11. 7 Tips to Secure a STEAM Internship This Summer

    Type
    Blog Post
    ... of internships as a result of the necessary restrictions on close personal contacts. However, that should...starting an application to assess your eligibility, interest, and career fit. Most internships share a common...likelihood of acceptance to the internship(s) you are interested in.     STEAMid is an app that helps you...Beyond the resume, a cover letter communicates your interest in a specific program, and institution. Consequently...paint a comprehensive picture of your academic interests, potential, and drive. Read the associated prompt...

  12. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...chemicals, hormones, proteins, etc., that you are interested in. The most obvious component of a biosensor... time. For example, if ATP was the molecule of interest (analyte), and accumulation of ATP activated GFP...for neuronal action potentials; it’s an ion of interest for many disciplines! Most of the sensors designed... the cytosol and sarcoplasmic reticulum are of interest to muscle biologists, while neuroscientists would... monitors associated with specific proteins of interest such as actinin. When tension is high vs. low ...

  13. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...permanently is to knock out the gene for your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need..., LoxP sites are knocked in flanking a gene of interest (or a single exon) and everything in between those...to a transcriptional repressor to your gene of interest. It will inhibit transcription instead of introducing...system is led, via a custom gRNA, to your gene of interest to temporarily increase expression. There are ...to for degradation. Since the tagged protein of interest is normally expressed, protein levels will quickly...

  14. RaPID Detection of RNA-protein Interactions

    Type
    Blog Post
    ...to identify proteins that bind an RNA motif of interest in living cells. Challenges with existing RNA-protein...processing. Require prior knowledge that the protein of interest interacts with RNA. Methods like cross-linking...followed by an antibody pulldown of a protein of interest, much like using ChIP for studying protein and...-binding proteins that interact with an RNA of interest. Current labeling methods are not ideal for in...of the bait, which for RaPID is an RNA motif of interest. See figure 1 below for details on the key components...

  15. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...approaching it. These include the traditional restriction enzyme/ligase-mediated method, the more recently...of clones that actually contain your insert of interest. Simply stated, ccdB makes cloning easier by selecting.../19 vectors and demonstrating the concept of restriction enzyme-mediated disruption of ccdB, leading to...replace ccdB with the investigator’s insert of interest. Hence correct clones are identified much more...swapping out” of ccdB with the investigator’s gene of interest.   CcdB resistant E. coli strains complete...

  16. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...Planarians are easy to buy, cultivate, and have interesting phenotypes to study. In addition, the Sánchez...biologists have to extract RNA from their organism of interest, use the RNA to create a complementary DNA (cDNA...for performing targeted knockdowns of genes of interest and was first discovered and characterized in ... other genes in planarians! For those who are interested in performing their own molecular cloning and...more curious” students on how to clone a gene of interest into the empty PR-T4P vector backbone. The empty...

  17. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ... a SmaI or XmaI restriction enzyme digest (the ITRs contain SmaI and XmaI restriction sites). Another ... transfer plasmid which encodes a transgene of interest that’s packaged into the virus. Two to five days...recombination of duplicated regions like the ITRs. Restriction digest screening for ITR deletion is still necessary...determine the optimal dose of AAV for my cells of interest. Every batch of AAV requires its own MOI optimization...

  18. PhD Applications After COVID

    Type
    Blog Post
    ...whose labs does research that aligns with your interests and goals.  Reaching Out to Potential PIs Even...comprehensive idea of that advisor’s research interests which can be helpful in creating your application...even better, one of their papers!) and express interest in their work Ask if they’re available for an ...professors will not only talk about their own interests, but will ask you questions about your research... yourself and learn more about your schools of interest. Benefits of Online and Hybrid Options Thanks ...

  19. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...gRNA(s) targeting a single genomic location of interest  When these three components are co-expressed...co-expressed, the gRNA targets the dCas9 to the loci of interest and dCas9 is biotinylated by BirA. From here, the...crosslinking captures chromatin interactions, a restriction enzyme digest cuts DNA into smaller pieces, and...line has to be created for each genomic loci of interest. This is cumbersome and also makes it hard to ...Cas9 Variants for Purifying Genomic Regions of Interest Here Brush Up on Your CRISPR Background with Addgene...

  20. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...often make cell lines that overexpress genes of interest; more recently we have also been using Addgene...co-transfected at a favorable ratio to the gene of interest (or, if your protein is fused with a fluorescent... different levels of expression of our gene of interest. Advantages of the agarose cloning method The...researchers, if growth properties in agarose are of interest, cell clones could potentially be picked following...Medical School in Baltimore. She is particularly interested in signaling peptide maturation in neural and...
Showing: 241 - 260 of 755 results