Skip to main content
Addgene

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

Information

In Category

We narrowed to 839 results for: LEA

Showing: 441 - 460 of 839 results

  1. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...also reversible: once the environmental trigger leaves, gene expression is no longer activated. However...N2A, 3T3) and organisms (Chavez et al., 2016). To learn more about the different types of Cas9 activators....3149 Additional Resources on the Addgene Blog Learn how to Choose the Best Cas9 Variant for Your Next...More about Genome-wide Screens with CRISPR/Cas9 Learn How CRISPR Has Been Used with AAV Resources on ...

  2. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...elements, insulators, and silencers; however, some "leakiness" of transcription may occur. This is normally ...small RNA expression shRNA From the human U6 small nuclear promoter Constitutive  Murine U6 is also used, ...lacI or lacIq). Can be induced by IPTG or lactose. Leaky promoter with somewhat weak expression. lacIq mutation...the Addgene Blog Browse All Plasmids 101 Posts Learn about Mammalian Vectors Read about Reporter Gene...

  3. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...protein is cleaved by proteases. Nowadays, with ample structural information available, it’s clear that the...protein is mislocalized. The APT1-mVenus fusion is clearly a better choice to work with. Inserting a fluorescent...the existing pool of protein within a cell and may lead to over-expression artifacts. This is an important...

  4. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    Published
    Sept. 5, 2017, 1:58 p.m.
    ... the nonhomologous end-joining (NHEJ) pathway, leading to nonspecific nucleotide insertions, dele­tions...allow blocking the PAM sequence (e.g. if it would lead to a missense or nonsense mutation), blocking the...the PAM as possible (see below to find out why). Please check our recent studies for details (Paquet, Kwart...28102837. Additional Resources on the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design ...

  5. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...degraded by uracil DNA glycosylase (UDG) and Exonuclease III (ExoIII) (step 4). UDG recognizes uracils...from newly synthesized DNA. Lack of both enzymes leads to a buildup of uracil and the insertion of uracil...limitation is sequencing depth and cost. You need at least 100-200 reads per variant, so the more codons you...to Generate Specific Mutations in Your Plasmids Learn How You Can Use CRISPR Pooled Libraries to Generate...

  6. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    Published
    July 10, 2018, 1:08 p.m.
    ...reproducibility. These alarming results are due, at least in part, to insufficient and non-standard record...exchange information regarding lab work with your colleagues, supervisors, clients or even students, your ...track and find data and information. If an employee leaves the institution, the transition is smooth as all...scinoteELN Additional Resources on the Addgene Blog Learn about more tools for reproducibility 5 great apps...

  7. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...Addgene blog Learn about research applications of antibodies in Introduction to Antibodies Learn more about...visualization methods. Stay tuned for a future blog post to learn more about these techniques!  Direct vs. indirect...what sort of protocol you will need to use. (To learn more about antibody validation, this commentary ...

  8. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ...Optogenetics Guide to learn more about how these modules work, and read on to learn how they are put to ...factors (GEFs) activate their partner Rho GTPase, leading to behaviors like lamellipodia or filopodia formation...these physical processes. Joachim Goedhart and colleagues in the Gadella Lab devised a set of optogenetic...

  9. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    Published
    April 8, 2025, 1:15 p.m.
    ...intuitive, qualitative interpretation of this gel is misleading. We can see why on the S-shaped curve. On the...on the band intensity axis. This effect causes misleading qualitative conclusions, and it’s also problematic...approaches Cartoons are nice and all, but we’ll leave you with this real-world comparison of different...of western blot normalization. May your blots be clear and your detection range be linear! Resources and...

  10. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    Published
    Nov. 25, 2015, 4 p.m.
    ...problems. This can lead to a lack of trust in the mentor and this training is clearly not equitable to ...assess the current state of research became more clear throughout the various sessions of the symposium...first couple of sessions of the symposium it was clear that, even though postdoc associations across different...

  11. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...strain-specific tropism.” For example, AAV-PHP.B doesn’t lead to enhanced transgene expression, compared to AAV9...mice use LY6A to cross the blood-brain barrier, leading to strain-specific tropism (See S3 table (Huang... can visit our Caltech Systemic Capsids page to learn more about that collection. Table 1: Systemic capsid...systemic AAV experiments and Addgene’s AAV Data Hub to learn more about the capsid you’re interested in. Then...Fish, K. N., & Pfenning, A. R. (2022). Machine learning sequence prioritization for cell type-specific...Troiano, E. C., Moghadaszadeh, B., Peacker, B. L., Leacock, K. A., Horwitz, N., Beggs, A. H., Wagers, A. J...

  12. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...expression, but causality and mechanisms remain unclear. Today, targeted epigenetic modification at specific...came epigenome-engineering tools - zinc finger nucleases and TALENs fused to epigenetic modifiers enabled...Christopher Newgard’s Lab has fused LSD1 to the smaller nuclease-dead Staphylococcus aureus Cas9 (Sa-Cas) and deposited...comments! Find Plasmids for Epigenetic Modification   Leah Schwiesow contributed to updating this article. ...

  13. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...multimer present in your sample. In these instances, please note that the supercoiled dimer band can sometimes... presence of a multimer in your sample remains unclear after visualizing it on a gel, one of the other...consistent with the size of a supercoiled plasmid dimer. Please note that it is common to see other plasmid confirmations...Ausubel FM. Recombination between bacterial plasmids leading to the formation of plasmid multimers. Cell. 1976...

  14. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    Published
    March 8, 2021, 2:41 p.m.
    ...reducing agent, typically containing thiols, to cleave disulfide bonds. Load the samples, one sample per...conditions from those used by the vendor then it may lead to nonspecific binding.  Whenever possible, include...in making your own knockout line for validation, please view this tutorial from Stuart Orkin's and Daniel...protein causes a thicker band, while less protein leads to a thinner band. It is important to note, however...

  15. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ... and the two HindIII digested ends will anneal leaving the 5’ end of your gene just downstream of the ...require you to visualize your DNA using UV light – please note that UV light can damage DNA and that proper... colonies on the backbone + insert plate (or at least more colonies than the backbone alone plate). Unsuccessful...Blog  Additional Resources on the Addgene Blog: Learn How to Verify Your Plasmid Try Out Golden Gate or...

  16. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...that introducing a Leu28Cys mutation in eDHFR does lead to covalent binding of the TMP-tag ligand (Gallagher...a different event where CP appears briefly then leaves. Images reused from (Ulrichs et al., 2023) under... if you are comfortable with it or can find a colleague to help you, synthesizing your own ligands can... D., Karassina, N., Zimprich, C., Wood, M. G., Learish, R., Ohana, R. F., Urh, M., Simpson, D., Mendez...

  17. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...Want to learn more about genome editing with CRISPRs? Check-out Addgene's CRISPR Resources Learn about...selective cassette minimizes the sequence added, leaving a 34 bp “scar”, but currently requires additional... editing and conditionally manipulate proteins. Learn more at his webpage or follow him on Twitter.   ...

  18. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...prevents both the endogenous RecBCD and SbcCD nucleases from digesting linear DNA introduced into the ...E. coli. Exo: Exo is a 5’→3’ dsDNA-dependent exonuclease.  Exo will degrade linear dsDNA starting from...with these promoters (lacI, araC, cI857) to limit leaky expression of the Red system.  Using a plasmid to...the red genes needed for recombineering, but also leads to expression of the int and xis genes which are...

  19. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ...epifluorescence microscopy wasn’t able to give a clear view of cellular movement in real time. Two-photon...microscopy has uncovered host-pathogen interactions leading to the understanding of effector function in infected...Weninger. "Dendritic cell behaviour in vivo: lessons learned from intravital two-photon microscopy." Immunology... Get Tips on Choosing Your Fluorescent Protein Learn Some of the Pitfalls of Working With Fluorescent...

  20. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    Published
    July 20, 2021, 1:15 p.m.
    ...such as fluorescent microscopy must be used. To learn more about the most suitable fluorescent microscopy...structure of the protein. In some cases this can lead to changes or a complete loss of protein function...more autofluorescence and nonspecific staining leading to false positives. Numerous dyes can be used for...Check out other methods using fluorescent proteins Learn more about antibodies Resources on Addgene.org ...
Showing: 441 - 460 of 839 results