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Showing: 181 - 200 of 1045 results

  1. The Golden Stopwatch Award: Recognizing Facilitators of Research

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    ...your TTO have a question? Refer them to Addgene’s Tech Transfer Team: techtransfer@addgene.org Congratulations...Addgene Help Center: Tech Transfer Technology Transfer Information Tech Transfer News: A Reintroduction ...Addgene account for Tech Transfer Offices. They’re packed with tech transfer resources (summaries, electronic...stats and strategies with tech transfer offices. Drop us a line at techtransfer@addgene.org. Albert Einstein...requests from Addgene MTA: material transfer agreement Technology Transfer: the exchange of intellectual property...addgene.org identifies the hold-up. Your Material Transfer Agreement is stuck in “pending.” Fear not, there... to make MTA turnaround faster with technology transfer offices (TTOs). The latest of these efforts is...

  2. Five Popular Model Organisms

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    Blog Post
    ...utilized mice to study how different mutations in leukemia impact different treatment regimens (Zuber ...example, the Stainier lab studied the proliferation and differentiation of β-cells in the the developing zebrafish...β-cells that originate in different regions of the pancreas and produce different levels of insulin (Hesselson...mouse (Mus musculus) Let’s start with the most preferred mammalian model organism of many researchers: ...injecting mice with retroviral vectors that harbored different mutations found in leukemia, the Lowe lab was ... gene expression in specific cells of the fly (Pfeiffer et al., 2010). Yeast (Saccharomyces cerevisiae...at a time. Zebrafish embryos are also laid and fertilized externally, allowing scientists to easily manipulate...

  3. Deep Dive: Statistical Tests (Comparisons)

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    ...test assesses the sum of differences between pairs). Samples arising from different subjects or populations...test assesses the difference of the group means). Unpaired t-tests can also be referred to as two-sample...significant differences between the means of your groups. However, it will only tell you if differences exist...samples are different from each other, or if every permutation of pairs within the five is different. To identify...groups to determine if they are statistically different. They are classic parametric tests and one of ... whether or not two samples are statistically different than one another, a two-tailed test should be ...control group. If you only care if one group is different in a certain direction (i.e., you just want to...

  4. Antibodies 101: Beyond Surface Labeling

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    Blog Post
    ..., Addgene uses 'labeling' to refer to antibodies and 'staining' to refer to small molecule dyes. Indirect...first fixed and then permeabilized, using two different buffer solutions. Fixing causes protein cross-linking...nucleus, a different fix/perm kit is required (to be precise, the fixing solution is different while the...plot (Figure 4b). Through this method, differences in proliferation between cell populations can be visualized...divisions).   Figure 4: Proliferation dyes can visualize proliferative capacity and speed of cells...lineage markers, like many of the cluster of differentiation (CD) proteins found on immune cells. However...use 'labeling' and 'staining' interchangeably to refer to any technique that combines a marker with a signal...

  5. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

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    ...maintaining the undifferentiated “stem” state and, Their potency or capability to differentiate into specialized...special types of cells that can develop or “differentiate” into more specialized cells with specific functions...function adequately [1]. Stem cells’ ability to differentiate into multiple cell types makes them useful models... cell has the ability to produce all of the differentiated cells in an organism, including the embryonic...Pluripotency: When a cell has the ability to differentiate into cells of any of the three germ layers: ...detail below [2]. Multipotency: When a cell can differentiate into multiple cell types of the same lineage...name suggests, cells under this category can differentiate into only one cell type. Muscle satellite cells...

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

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    Blog Post
    ... virus can be stored at 4 °C and buffer exchanged the next day. Refer to the AAV Iodixanol Gradient Ultracentrifugation... AAV packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid...help mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene of interest ...more details. Pro tips I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple...to grow multiple bacterial cultures of your AAV transfer plasmid at 30 °C instead of 37 °C and then screen...restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, like NEB Stable...

  7. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

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    ...created phages. Transfer of this DNA to another cell is referred to as transduction. Transferred DNA once inside...donor can transfer genetic material to the recipient bacterium. The genetic material transferred is commonly...Horizontal gene transfer (HGT) is the movement of genetic material between organisms. It plays a key ...unique to the lab. Let’s take a look at these different methods of DNA insertion. Transformation Transformation...Various factors promote natural transformation in different bacteria such as growth phase of the cells (Baltrus...plasmid, the genes on the plasmid can give them different phenotypes. For example, this plasmid encodes ...cell. Once inside, phages can follow one of two different life cycles: lytic or lysogenic. Lytic phages ...

  8. Viral Vectors 101: Gamma-Retroviral Packaging Systems

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    ...pseudotype γ-retrovirus using different envelopes to modify tropism. Briefly, different envelope plasmids can ...engineered γ-retroviruses differ from lentiviruses? There are a few important differences between engineered ...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...Vectors 101 eBook, 1st edition.  References and resources References Swift S, Lorens J, Achacoso P, Nolan...following components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA ...sequences, which facilitate integration of the transfer plasmid sequences into the host genome. Typically... the host genome upon viral transduction. Most transfer plasmids available from Addgene were derived from...

  9. What's the Best Way to Elute and Store Your Plasmid DNA?

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    ...Tris-EDTA (TE), just Tris buffer, or some other variations. Does it make a difference? Short answer: yes. This...choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose could...TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving the stability of your preparation...can be Tris buffer (with no EDTA) or water. These are two of the choices that companies offer for the elution...preparation can be eluted in water or in your buffer of preference, and it will be stable for years. Plasmid... preparation for a long time. Tris buffer controls the pH, while the EDTA chelates any divalent cations...through their kit. The advantage of using a Tris buffer is that it allows better control of the pH preparation...

  10. Plasmids 101: The Wide World of Natural Plasmids

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    ...through horizontal (or “lateral”) gene transfer. Horizontal gene transfer can occur naturally through transformation...conjugation (direct transfer from one cell to another). Bacteria need special fertility genes to be capable...Emily P. Bentley in October 2024. References and Resources References Demanèche, S., Bertolla, F., Buret...Here at Addgene, we often refer to plasmids as lab or experimental tools. They certainly are very handy... evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al., 2001) Beneficial...humans only noticed last century! Nevertheless, the transfer of plasmids carrying these genes between bacteria...Historically, colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.” Virulence...

  11. Plasmids 101: Broad Host Range Plasmids

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    ...host range is determined by the different hosts that a plasmid can transfer to, replicate in, and persist...stable, can be easily cloned in different bacteria, and can be transferred to various experimental hosts... that can replicate in different bacteria and express proteins within different host organisms. pBBR1-...recombinant vectors at Addgene!   References and resources References Doran, K. S., Konieczny, I., & Helinski...host range, identify a few of the features that confer broad host range in plasmids in nature, and provide... they carry among organisms belonging to many different phylogenetic groups (Jain and Srivastava 2013)...be used by scientists across disciplines.  What confers broad host range in plasmids? Not all plasmids ...

  12. A Guide to Designing a Scientific Poster: Content, Layout, and More

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    ...your poster Check conference guidelines To begin, you should always follow conference rules when designing...useful to use two different font styles; one for your section titles, and a different one for the body ...distinguished. The UCLA Library offers some Powerpoint based templates with different column numbers for a good... the most of your conference attendance Read all of our blog posts about conferences .... But, before you can show off your work at a conference, you need to first make your poster. While a ...understand that presenting your work in poster format differs in many ways from writing a manuscript. If you ...you have ever attended a scientific conference, or even a lab recruiting session at your university, you...

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

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    ...not transfer precisely evenly across the entire blot. So how can you accurately compare different lanes...assuming that all differences in the loading control are due to loading and transfer errors — not to any...reflect massive differences in protein abundance. Validating the assumptions for different loading controls...detection range be linear! Resources and References References Aldridge, G. M., Podrebarac, D. M., Greenough...errors like uneven pipetting or irregular membrane transfer. Figure 1: A loading control is a type ...Researchers use later steps in this chain (blue) to infer or calculate things about earlier ones. In practice...relationship poses problems for proteins of very different abundance, consider the following example gel....

  14. The MTA at Addgene

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    ...There are many different flavors of MTAs, but the Uniform Biological Material Transfer Agreement (UBMTA...process that differs.  When you deposit a plasmid with Addgene, your organization’s tech transfer office is...for more information. Happy transferring!    Many thanks to our Tech Transfer team members Maura Coughlin...Addgene, then you’ve been involved in the material transfer agreement (MTA) process. Even though this process...curious about the hows and whys of the material transfer agreement, read on!  What is the MTA?  An MTA...UBMTA) is one of the most commonly used ones for transfers between academic institutions.  What is the UBMTA...the creation of a standardized agreement on the transfer of biological materials in 1995: the UBMTA. Because...

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

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    ... deposited by Promega here.    References and resources   References Allikalt, A., Purkayastha, N., Flad... scientists. Seeing inside these specimens has offered illumination on biochemical processes that are ...Figure created with biorender.com. Fluorescence refers to the emission of light by a substance (fluorophore...energy due to non-radiative energy losses. This difference in emission results in a slightly longer wavelength...phenomenon is known as the Stokes Shift, or the difference in wavelength between peak excitation spectrum...vary in their chemical structure, leading to differences in their excitation and emission wavelengths,...target specific organelles like mitochondria, and different dyes can result in varying cytoplasmic background...

  16. Luminescent Imaging with Nano-lanterns

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    ...Renilla luciferase variant fused to an FP; when supplied with a luciferase substrate, the luciferase transfers...bioluminescence resonance energy transfer (BRET). Photons emitted by a Renilla luciferase variant are used to excite...problems of low luminescence By itself, Renilla luciferase has a low quantum yield, and when transiently...Nano-lantern, a fusion of a mutagenized Renilla luciferase (RLuc8) with Venus. This luminescent technology...need for an excitation light source. Instead, a luciferase substrate like coelenterazine is supplied in ...technique’s versatility and adaptability to many different contexts. Improving Nano-lanterns In 2015, Takai...eNL constructs use the brightest characterized luciferase, NanoLuc, and its substrate furimazine. In addition...

  17. Understanding GPCR Signaling Using TRUPATH

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    Blog Post
    ...Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which... acceptor fluorophore. The difference for BRET is that the energy transfer is between a luminescent donor...number of different experimental approaches. They first assessed the activation of different members of...response to different agonists. These studies showed the promiscuity of GPCRs for different G protein transducers...neurotransmission. But with approximately 950 different human genes encoding GPCRs (Takeda S et al., 2002...a vast number of signaling cascades, inducing different cellular responses. One of the biggest advantages...measure the physical association of combinations of different Gα, Gβ, and Gγ subunits. When inactive, these ...

  18. 27 Hot Plasmids from 2016

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    ...control many cellular processes such as differentiation, proliferation, and apoptosis. In humans, mutations...Protein A-Renilla luciferase and the “prey” proteins are tagged with Firefly luciferase. The bait and prey...variable availability of enhancer sequences in different chromatin states. Alexander Stark and colleagues... were deep sequenced to identify the gRNAs that confer protection to WNV. From this screen the authors...process of cloning, combining, and testing these different parts can be quite arduous. The cloning process...consists of a variety of plasmids containing many different promoters, RBS’, coding sequences (CDS), and terminators...To facilitate innovative cancer research, the Reference Reagents Group of the NCI RAS Initiative, led ...

  19. Viral Vectors 101: AAV Variables That Matter

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    ... variables.    Serotypes Serotype refers to the different protein compositions of AAV capsids, which... a virus enters the cell. Different serotypes utilize a number of different receptors, co-receptors, and...burdensome in the brain where many different cell types, or even different phenotypes of the same cell type...working in several different systems or want to transduce a large number of different cells. Promoters ...doi.org/10.3389/fnmol.2014.00076 References and Resources References Aschauer, D. F., Kreuz, S., & Rumpel...validation and optimization. Although there are different ways to ultimately achieve success, we find it...validation first.  AAV-mediated gene expression can differ extensively depending on the experimental system...

  20. Antibodies 101: Immunoprecipitation

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    ... However, the two different approaches allow for optimized strategies for differing amounts of target ...urea-containing lysis buffer. Alternatively, the protein can also be eluted using a low pH buffer such as 0.1M ...for other antibody topics.     References and resources References:  Björck L, Kronvall G (1984) Purification... appropriate for most situations.  A slightly different method is desirable if the antibody is expected...Immobilization is the key to immunoprecipitation and simply refers to the process of anchoring an antibody, often ...eliminating the risk of the primary antibody interfering with downstream assays. This is particularly ...than those of agarose beads, despite the size difference. Magnetic beads are particularly advantageous...
Showing: 181 - 200 of 1045 results