Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 789 results for: mpl

Showing: 261 - 280 of 789 results

  1. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    Published
    March 21, 2024, 1:15 p.m.
    ... of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes, and...attached to self-labeling proteins. Some common examples of these self-labeling proteins would be SNAP-...of protein substrates/peptides and antibodies. Example applications of this include coupling to an antibody...photons absorbed over the photons emitted. For example, DAPI is much dimmer than Alexa Fluor 488, and ...cells without causing undue toxicity. A prominent example is DAPI, a dye that binds specifically to the AT-rich...fluorescence-based techniques in biotechnology. Recent examples of advancements include the Janelia Fluor(R) (...finding the right dye for your experiment more complicated. We hope this post helps you in your search for...

  2. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    Published
    Oct. 20, 2022, 1:15 p.m.
    ...the RNA from degradation, several simple methods have been employed which modify the sugar or phosphate... when scientists talk about their precious RNA samples. RNA is inherently less stable than most macromolecules...suggest coupling several of them together, for example phosphorothioates with 2’-O-methyl modifications...make your gRNA more efficient on a budget is to simply swap out several of the ribonucleotides for deoxynucleotides...RNA to control the entire editing system with a simple LED light. Lighting up your gRNA with fluorescent...

  3. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ... on excitation power in a linear fashion. This implies that reducing the power 2-fold does not reduce ...fluorescent proteins can be directly compared. An example of a photostability measurement of various cyan...results in a loss of fluorescence, which, in its simplest form, can be described by a mono-exponential decay...intensity decay of a fluorophore does not follow a simple mono-exponential decay (Shaner et al, 2008; Bindels...applications in which far less power is used. For example you might find that you can get useful data from...fluorescent protein. In figure 1 we provide an example of such a comparison between cyan fluorescent proteins...

  4. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ... The destination vector contains sites with complementary overhangs that direct assembly of the final ...plasmid (Option 1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning... reaction. For detailed Golden Gate protocols, complete with helpful tips and tricks, see The Sainsbury...is to "domesticate" your fragment: PCR-based amplification can be used to create silent point mutations...system is an incredibly powerful tool for cloning complicated constructs in a single, high-efficiency step....step. References: Sequential amplification of cloned DNA as tandem multimers using class-IIS restriction...

  5. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ... or Cas nickase)  a guide RNA that targets the complex of TadA and dCas to the genomic DNA of interest...effect of removing the wild-type TadA from the complex the researchers developed a second set: The ABE8... the team replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes... ability to use non-standard Cas9 in the ABE8 complexes enables a broader targeting scope for base editing... the corrected cells back to the patients. For example, defects in hemoglobin (𝛽-globin), which cause...that require high DNA editing specificity, for example for potential therapeutic approaches, the authors...

  6. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...plasmid are not always the best for others. For example, in our hands we find that a double harvest approach...larger genes such as Cas9. See figure 1 for an example of how changing transfection ratios can affect ... For some transgenes, transfection optimization simply is not enough to get a sufficient titer; this is... volume than with the unconcentrated prep. For example, at Addgene we typically concentrate 100-fold meaning...volumes of lentivirus routinely, you may opt for simply increasing the number of 10 cm2 dishes. While this...available, cost and downstream applications. For example, while the PEG precipitation method is relatively...

  7. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ... features of the gene intact. Here, we show an example gene targeted for tagging with GFP at the C terminus...interest. For a C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the ...percent of cases, the stem cell will seek out a template for the repair process. By flooding the cell with...using the introduced donor plasmid as a repair template. This results in the insertion of the GFP tag ...tag precisely where we need it, in this example, at the C-terminus of the gene of interest. To get the molecular...CRISPR/Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and the donor plasmid into the cells we use electroporation...

  8. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    Published
    Dec. 1, 2015, 3:30 p.m.
    ...the ribosome complex and Dr. Steitz quipped that RNA in the ribosome is in a complicated conformation ...Lowe lab have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs...long hypothesized that life started with RNA and a simple cell membrane. His recent work shows that they ...replication with no proteins). The chemistry is complex and this work requires an interdisciplinary team... they do it. Of course, No conference would be complete without CRISPR Michael Terns from of the University...

  9. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    Published
    July 23, 2015, 2:30 p.m.
    ...are often thought of as simple organisms , they have many levels of complexity, especially when trying...learning how to grow and care for the worms. As an example of one of the problems we work collaboratively ...We attempt to deconstruct and understand this complexity using existing C. elegans mutants. For instance...data is freely available, and the collection of samples and sequencing is being organized and funded by...Kittybiome project gives them access to rich and complex microbiome data for analysis. And, well, cats.....

  10. Transferable Skills: Negotiation

    Type
    Blog Post
    Published
    March 3, 2022, 2 p.m.
    ...though three examples depend on many specific details about the individuals, each example provides a scenario...machine? Let’s walk through a more specific example. Perhaps you and your lab mate are negotiating ... something valuable. Importantly, as the third example shows, the ultimate resolution can involve ideas...your organization. In my non-academic roles, some examples of negotiation have been: A job offer conflicts...explore while also ensuring that my work gets completed? I want my company to pay for me to attend a conference...

  11. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...realized how perfectly flat the complex is on this surface”….where the complex is docked on the membrane of...like “Oh,…I didn’t quite appreciate that….” For example, when Tom Steitz was presented with a physical ... close to its reported cleavage site on the complementary DNA strand. And this is why we love working ...cleavage”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting...double-stranded DNA, recognize a PAM sequence, allow the complementary base pairing between the guide RNA and the target...

  12. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ...endogenously expressed SCF complex members, an E3 ubiquitin ligase complex. Finally, when auxin is added...fusion protein with the endogenous CRBN E3 ligase complex which will polyubiquitinate the fusion protein....target protein-of-interest and the CRBN E3 ligase complex leading to ubiquitination and proteasomal degradation...acid (IAA), the TIR1 F-box associated E3 ligase complex will ubiquitinate the AID-tagged protein which ...Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc...

  13. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ... millions of people die from cancer-related complications each year. Despite these grim statistics, new...cancer, really? While cancer is extraordinarily complex at the molecular level, at the cellular level there...cancer research has focused on picking apart the complex cellular pathways involved in the development of...pathway. Its major appeal is that it enables the simple construction of vectors to study the function of...transcript most commonly found across all TCGA tumor samples is represented, making the results of any such ...biggest challenges facing researchers today is the complexity of the pathways involved in the development of...

  14. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...allowing very low background level of fluorescence. Examples for such photoactivatable or photoconvertible ... and their emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent...sensor), and iGluSnFr (glutamate sensor). More examples of these biosensors can be found at Addgene. ...also is also called Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing...system can excite, and detect the emission. For example, if your microscope has only two lasers, at 488nm...minutes after it is translated to a few hours. For example, superfolder GFP (sfGFP) and mNeonGFP can fold ...

  15. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ... takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes. ... centrifugation. At this point, the plasmid DNA sample can be treated with RNase to remove RNA contamination...directly into a PCR buffer, where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based ...waste footprint. The Zhao and Deng labs developed a simple method for regenerating silica columns that come... reduce the number of spin columns you need to complete your research. Do you know of other nucleic acid...

  16. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    Published
    Feb. 6, 2020, 3 p.m.
    ...major findings and then focusing more on broader implications rather than diving deep into the individual ...carefully chosen for each group and topic. For example, it may be useful to adapt the second version (...stay abreast of current trends in the literature, sampling a breadth of work from a field (perhaps even two...to allow those who could not attend a meeting to simply stay caught up and to allow those who may be interested...get feedback from those who didn’t attend. For example, we got a good bit of early feedback from those...own knowledge of the field. Perhaps interest has simply waned. When attendance slumps below a certain level...

  17. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...mediate cleavage rather than the multi-subunit complex employed by class 1 systems. A putative new class 2...nucleases. Using Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover...than following it. The Cpf1 crRNA is also much simpler in structure than Cas9’s; only a short stem-loop...CRISPR systems based on Cas9 were thought to be the simplest CRISPR systems and the easiest to adapt to genome...confirmed. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Comparing Cpf1...

  18. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...Its RNA-guided nature makes CRISPRa much simpler to implement in genome-wide screens over cDNA libraries... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating... must be screened to identify effective ones. Simplified in vivo delivery using SPH mice CRISPRa has largely...population, as expected. Although this strategy was employed to study neuronal differentiation, it could also...

  19. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    Published
    Aug. 14, 2018, 12:42 p.m.
    ... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...photostable than GFP. In addition, antibody-based amplification of their signals is weak and increases background...sites for primary antibodies which can be used to amplify the direct smFP signal within sub-cellular structures...used to distinguish highly similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell...super-resolution STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes...smFPs retain their ability to be detected after sample preparation better than standard proteins tags ...

  20. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...of the viral capsid proteins VP1, VP2, and VP3. Samples of viral preparations are subjected to polyacrylamide...proteins as a fraction of total protein present in the sample is also determined and used to determine purity...shown relative to the total protein present in the sample.   Sterility Viral vector preparations are ...rigorous barcode matching system that follows the sample from the DNA tube all the way to the cleared viral...throughout production, ensuring the identity of each sample from the DNA tube all the way to the viral preparation...Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers ...
Showing: 261 - 280 of 789 results