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We narrowed to 520 results for: HA tag

Showing: 301 - 320 of 520 results

  1. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    ...DRSC/TRiP- Functional Genomics Resources website has in vitro reagents and tools to help you knock down...tools. The Vienna Drosophila Resource Center also has collections of flies for in vivo gene knock down ...finishing his immunology PhD at Harvard University. He has had nearly a decade of experience working with Drosophila...three larval stages where larvae molt in order to continue growing in the next larval stage. After the ...the third larval stage, Drosophila will pupate, which takes about one week from the initial deposition of...through their translucent bodies. This is the ideal stage to select for females because you can be certain...

  2. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...CRISPR/Cas9 technology has been a breakthrough for the genome-editing field. It has the possibility to treat... applications.” Li’s team noted that CRISPR/Cas9 has been used to generate mutations in a wide range of... expanded organoids. “CRISPR in adult stem cells has huge potential, because the genomes of adult stem...of the cystic fibrosis report. “This is a big advantage over induced pluripotent stem cells, which need...

  3. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ... your research. RNA editing advantages RNA editing has multiple advantages over more traditional DNA editing...discussed above, the ability to edit RNA has multiple advantages, including reversibility and use in non-dividing...will preferentially edit an adenine if the template has a cytosine mismatch at that position. PspCas13b-...cytosine deaminase. Three rounds of rational mutagenesis on the ADAR2DD residues that contact the RNA ...RNA-sequencing, the authors performed rational mutagenesis and identified that a S375A mutation in RESCUE...

  4. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...Cpf1-driven systems has added another option to the CRISPR toolbox. Cpf1’s staggered cleavage pattern opens...frequency of desired HDR edits, but the possibility has not yet been experimentally confirmed. To prevent...adaptation of Cas13 to RNA targeting and RNA editing, has further shown the diversity of CRISPR-based systems...respectively), both cleave in a staggered pattern similar to FnCpf1. Potential advantages of Cpf1 over Cas9 Type...HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA rather than ...nt upstream of the PAM site, Cpf1 cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18...

  5. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...2018, uses quadrants to gate and has numbers representing the percentage of cells in each gate. Used under...right corner.      Figure 2: This flow plot has two populations that are B+ and one population that...Figure 3 above, until the population of interest has been successfully identified and isolated.  Following...2017). Neonatal immune response to rhinovirus A16 has diminished dendritic cell function and increased ...numbers by gated populations; this indicates the percentage of total events contained in the gated population...

  6. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    ...and post-docs alike are wondering where the time has gone. Are we any closer to graduating, publishing...unsure of the next steps. Harvard PI Angela DePace has put personal development at the center of her lab... and postdoc training. On the other side, the PI has an opportunity to check in with lab members, determine...technique are beneficial for researchers at all stages. Rather than succumbing to academic inertia, it...

  7. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ... and software development has caught up with it. The peer review process has been challenged with many...targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable...needs of each individual; the  community as a whole has yet to settle on any true kind of value. I’ve dubbed... genome editing tool, the CRISPR/Cas9 technology has been surrounded by a whirlwind of research activity...significant improvement. As a result, the software piñata has been filled with slight variations on a theme without...evaluate off-targeting, but currently no software has been designed for this purpose. This second area ...findings to help everyone else. CRISPR technology has reached into so many different communities that it...

  8. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...et al., 2016; Duret et al., 2019). But the field has sometimes seen controversy, and the interactions ...yet to be demonstrated (or invented), its behavior has already been successfully reproduced by several other...tools. And, since MagLOV’s parent protein AsLOV2 has already been adapted into a variety of optogenetic...  Figure 1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT magnet...effect (many do). Fusing EGFP to the self-labeling FlavinTag made a reliable one-component magnetoresponsive... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants...significant gains! After a few more rounds of mutagenesis, they eventually found a version of AsLOV2 with...

  9. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...the State Museum has been the best supporter and most productive partnership CINSO has had. Volunteer! ...provide online. Twitter is an excellent resource that has gained a lot of traction with researchers. It is ....  Try new things to further your career Take advantage of opportunities provided by your programs and...including workshops and networking events. I took advantage of as many of these resources as I could. Networking...front work was done for me. I just had to take advantage of it. Blog and tweet I started my blog, Born ... been “traditional,” I was able to fully take advantage of my graduate experience. Melanie Fox is currently...

  10. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...cleavage with some of these peptides, no consensus has been reached on which peptide works best. Likely ... is easiest to start with a plasmid that already has the multicistronic element and reporter cloned in... unique genes, you can start with a plasmid that has multiple cloning sites flanking the multicistronic...expressed from the same cassette is sometimes advantageous, particularly when only a portion of the plasmid... mRNA. However, some bicistronic vectors take advantage of an element called an Internal Ribosome Entry...bicistronic vectors; however, they do have some disadvantages. These elements are quite large (500-600 bp)...vector.3  2A Peptides To overcome some of the disadvantages of the IRES element, scientists have adapted...

  11. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...target in cis and has no trans activity. AsCas12a, on the other hand, generates staggered DNA nicks at its...targeting. Fast forward a decade, and the CRISPR field has exploded with dozens of Cas enzymes and variants ...scientific community. What are the features? CasPEDIA has collected relevant information on all the Cas enzymes...learn about Cas domains. CasPEDIA either directly has this information or links out to this content and...

  12. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...Counter-selection with sacB has revolutionized allelic exchange. Even now, only half our lab has switched over to...to pTOX. However, we feel it has clear advantages, and are hopeful that continued development of allelic..., 2014). In recent years, CRISPR-Cas9 technology has also aided in the creation of scarless mutations,...isolated are those where the allelic exchange vector has undergone single crossover and integrated into the...the lab of Matthew Waldor at Harvard. Now that he has an efficient allelic exchange system, he is so excited...

  13. Evolution of Lab Techniques

    Type
    Blog Post
    ...amount of high-quality DNA, real-time PCR has several advantages including easier automation, higher throughput...abundance. If a scientist wants to know if an organism has a particular gene mutation, southern blotting can... DNA is complementary to the probe (for example, has the sequence of the mutated gene the researcher is..., two DNA fragments are observed because the DNA has the site where MstII cuts. In a patient with sickle...DNA fragment is observed. Today Southern blotting has been largely replaced by real-time PCR to answer ...questions (see below). Importantly, although their role has changed, restriction enzymes are certainly not dead...

  14. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...refined to create Brainbow 3.2. Brainbow labeling has also been applied to cell marking and lineage tracing...example, Brainbow-1.0 expresses RFP when the construct has not undergone recombination. This default state was...neuronal pThy1-Brainbow constructs, Addgene also has two Brainbow adeno-associated viral vectors (AAV)...Drosophila and zebrafish. Brainbow in Drosophila has aided in the mapping of neural circuits, such as ...neuromuscular junction. In zebrafish, the method has become very useful in lineage tracing; “Zebrabow”...Lichtman estimate that Brainbow’s colorful labeling has decreased the mapping time for a given section of...AAV) available - AAV-EF1a-BbChT and AAV-EF1a-BbTagBY. These constructs contain two XFPs each due to size...

  15. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...Synthego’s blog. CRISPR-based genome engineering has revolutionized the gene editing field by making experimental...often under-appreciated or over-looked.  Synthego has developed a new tool called ICE (Inference of CRISPR...increased flexibility and scalability, the ICE software has two analysis formats: sample by sample analysis, ...comparable (within r2=0.96) to that of NGS data. Advantages of ICE Determine Editing Efficiency and Types...of CRISPR editing. ICE software identifies the percentage of genomes that have been successfully modified...(SpCas9). ICE Score - The editing efficiency (percentage of the pool with non-wild type sequence) as determined...size of the insertion or deletion along with the percentage of genomes that contain the indel. The discordance...

  16. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ... animal moving around to explore the environment has a different motivation (and brain areas associated... certain forms of drug addiction. When an animal has been exposed to morphine or other stimulating drugs...involved in locomotor sensitization? Previous research has shown that regions such as the nucleus accumbens ...dark compartment (Howerton at al. 2014). Addgene has several recombinase-expressing ready-made AAV vectors... tests.    Forced swim test The forced swim test has been used since the 1970s to test the efficacy of...rather than being out in the open (risky). Taking advantage of these characteristic traits, rodents can be...

  17. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...generally mediates broad cell tropism and has a high percentage of neuronal targeting after stereotaxic ... damage to the brain tissue. The injection speed has to be slow to allow the viral particles to diffuse...order to minimize the pressure into the tissue, but has to be large enough to target larger areas, typically...Alternatively, for larger brain areas, repeated staggered injections may be considered. Timing: High expression...

  18. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

    Type
    Blog Post
    ...antibody is biotin-conjugated, streptavidin (which has a high affinity for binding biotin), is conjugated...detecting one protein from the sample. A direct ELISA has many fewer steps than a sandwich or competitive ELISA...linear portion or the OD of your experimental samples has reached 4.0, you can resolve this issue when you ...on the Addgene blog Learn about the advantages and disadvantages of using monoclonal antibodies. Read...

  19. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    ...beginning to see Addgene as a large data set. Addgene has over 65,000 plasmids in the repository, each verified...predictors because they are just that: rare. “Nature has many genes where there are not rare codons and many...at the time, got involved. Along with Philipp Pfingstag, a visiting graduate student from TU München at...intrinsic properties (GC content or rare codon percentage) and comparative properties (percent sequence...algorithm, they found that  GC content and rare codon percentage were not good predictors. There is not much variation...  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil C. Thompson. "Gene synthesis allows ...

  20. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ... and serotype, respectively. For example, AAV2/8 has a type 2 ITR and a type 8 serotype. Almost all vectors... components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that your gene...usually quite stable and in some mammalian tissues has shown expression for several years (Wonjo et al.,...scAAV vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) ...specific cells and/or tissues is one of its biggest advantages. Specificity can be achieved using specialized...
Showing: 301 - 320 of 520 results