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We narrowed to 866 results for: FER

Showing: 321 - 360 of 866 results

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

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...a property of the fluorescent protein. This can differ between fluorescent proteins in the same spectral...dependence on excitation power is important since different fluorescence imaging strategies use widely varying..., TIRF or spinning disk confocal use entirely different excitation power regimes. Therefore, it is unpredictable...the best way to compare the photostability of different fluorescent proteins? Measuring photostability...excitation power. By repeating these measurements for different fluorescent proteins and by comparing how the ...measurements reported in the literature are performed in different ways. We describe a number of issues with some...

  2. Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing

    Type
    Blog Post
    Published
    May 5, 2015, 3:54 p.m.
    ...scientific manuscript will require numerous other references to scientific literature to substantiate the ...literature related to the topic. Finding reliable references and sorting out what they mean is no small task...Did they perform all of the appropriate controls? Refer back to the methods section for finer points of ...inspiration and accountability. There are a few different ways to automate the process as well. Sign up ...with scribbled notations in the margins. If you prefer, printing and hand-writing notes is still possible... Check out Papers and Mendeley – these programs offer more options for searching and sorting your own ...

  3. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ...such as p300 and CBP histone acetyltransferases or CARM1 methyltransferase. This attendant signaling feature...years evolved from chloramphenicol acetyltransferase to luciferase, the basic assay remains widely used...distinct complexes may explain how more than 300 different coactivator proteins individually contribute to...patterns of nuclear receptors across multiple different tissues in normal and disease states, their circadian...

  4. Lab Automation at Addgene

    Type
    Blog Post
    Published
    March 22, 2018, 1:28 p.m.
    ... one. As a starting point, I would recommend conferences like Lab Robotic Interest Group (LRIG). There...customizability. It has a large deck that can house many different types of devices. As long as a device fits on ... - one pod can pick up 96 tips for easy sample transfer between 96 well plates. The second pod can pick...out your samples. This can be useful if you’re transferring samples from 96 wells plates onto agar q-trays...follows all the same steps and uses the similar buffers as would be used for a mini prep, except it does...video library Check out our molecular biology reference pages ...

  5. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    Published
    July 14, 2016, 2:30 p.m.
    ...some of Cas9’s shortcomings. Cpf1’s preferred PAM is 5’-TTN, differing from that of Cas9 (3’-NGG) in both...pattern opens up the possibility of directional gene transfer, analogous to traditional restriction enzyme cloning... cloning. Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells...immunogenicity and range of serotypes allowing preferential infection of certain tissues. However, these...Cpf1 fits into the genome editing landscape. References 1. Kim D, Kim J, Hur JK, Been KW, Yoon SH, Kim...

  6. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...plasmids for Borrelia burgdorferi from Christine Jacobs-Wagner’s lab produce different levels of dCas9 expression...system developed to track cell-cell interactions By Jennifer Tsang GFP-based Touching Nexus (G-baToN) is a ...cell surface. When the cells interact, GFP is transferred to the receiver cell, and the receiver cell turns.../genetics/iyab006       The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR...expression and contain different antibiotic resistance markers. New CRISPR pooled library from Keith Joung...

  7. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...focus on gene editing techniques. References and resources References Akhtar, W., De Jong, J., Pindyurin...means we could design hundreds to thousands of different pegRNAs for each desired edit, it simply becomes...many more. Once we had our datasets with all the different features and editing efficiencies, we could train...offline via GitHub. The model provides users with different designs of pegRNAs, ranked by their PRIDICT/PRIDICT2.0... predictive power is sometimes limited across different targets due to variations in chromatin context...

  8. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    Published
    April 18, 2024, 1:15 p.m.
    ...we used different categories (Figure 2C). Note that in Figure 2, the numbers on the X axis refer to distribution...previous Addgene CRISPR publications.  References and resources References Pyhtila, B., Kasowitz, S., Leeson...through the years.  Libraries are divided into different categories based on function: knockout (KO), activate...have CRISPR plasmids deposited from labs in ~40 different countries, and we’ve shipped CRISPR plasmids to...

  9. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...but timer activity can be calibrated to these different conditions, enabling the use of a given timer ... of Neurog3 (a gene controlling pancreatic differentiation) in mouse embryos (Miyatsuka et al.) Distinguish... Let us know in the comments section below! References 1. Terskikh, Alexey, et al. ""Fluorescent timer...11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine...and Michael S. German. "Chronology of islet differentiation revealed by temporal cell labeling." Diabetes...programmable synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-...

  10. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ... Snapgene and Geneious software. References and resources References Chadeuf G, Ciron C, Moullier P, Salvetti...sequencing reads (~150 bp each) are aligned to the reference sequence of the plasmid used to create the viral...more than 90% of the reads should align to this reference sequence. Second, we perform a megaBLAST search...search on all the reads that did not map to the reference sequence. The resulting list of hits is then carefully...reads into longer DNA contigs without using a reference sequence. The five contigs with the most total...

  11. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ... the protein within a living cell. Analyzing differences in protein localization and transport between...signals not visible to the human eye and are indifferent to subjective color intensity perception. Open...statistics that can be used to compare between different experimental setups. A colocalization analysis...one fluorescent signal in a channel setup for a different fluorescent signal) and photobleaching i.e. FP...molecules more closely is Förster Resonance Energy Transfer, FRET. Additional opportunities: Tracking intracellular...Markers of Subcellular Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM...

  12. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...subtypes of A–G, which come from different species and will have different compositions of Cascade. Although...degradative power in the future! References and Resources References Loeff, L., Brouns, S. J., & Joo, ... archaea. Class 1 is further broken down into different types, with Cas3 systems making up type I, the...effector. Type I has subtypes A through G, with different compositions of the Cascade complex. Created with...

  13. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...chromoprotein and how is it different from fluorescent proteins? Before going into the difference between chromoproteins...research Read our Molecular Biology Reference Find plasmids for FRET References Blanquet RS, Phelan MA (1987...imaging microscopy-based Förster resonance energy transfer (FLIM-FRET) in living cells (Figure 4). They created... Jennifer Tsang...

  14. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...rely on two-step hierarchical approaches using different Type IIS restriction enzyme specificities at each...Factors such as enzyme efficiency, stability, and buffer compatibility have placed practical limits on single...Assembly test systems Our research focused on three different levels of assembly: High efficiency and accurate...biotechnological cloning efforts for over 50 years, prefers ligation of Watson-Crick base pair substrates, ...products to market, New England Biolabs, Inc.       References 1. Vladimir, Potapov, et al. "Optimization of...cloning protocols Check out our molecular biology reference pages ...

  15. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...be present in a single lentiviral transfer vector or separate transfer vectors. May contain reporter gene...outcomes can be most effectively achieved with different CRISPR components. Select the appropriate CRISPR...4.5kb). CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles. Transient...cell lines is quite broad and includes several different methods, so we've summarized some key features...should review the current literature about their preferred model.  If your expression system is not well ...

  16. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    Published
    Aug. 30, 2022, 1:15 p.m.
    ... as fixing and permeabilizing.  References and resources References Brown-Harding H (2020) Fixation artifacts...Formalin: You may also come across formalin, which refers to a saturated solution of formaldehyde (saturated...100, Tween-20, or saponin. Each detergent has different chemical properties that will impact the level...concentration.      Figure 2: Examples of different permeabilization strategies and when to use them...fixation; they require an additional step generally referred to as Antigen Retrieval. I chose not to get into...

  17. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ... a nutshell, is the systematic combination of different genetic sequences into plasmid DNA. In high throughput... specific sequences of the genetic elements may differ (e.g., a set of various mammalian promoters), the...cloning can be used to combine the functions of different genetic elements to generate non-natural tools...assembling these modular types of constructs, and different methods are suited for particular downstream applications...Cloning Strategies Method Technology Pros Cons References Golden Gate Type II restriction enzymes Can...Kits at Addgene Find Plasmid Cloning Protocols References 1. Anderson JC, Dueber JE, Leguia M, Wu GC, Goler...

  18. Getting to Know Expansion Microscopy

    Type
    Blog Post
    Published
    June 20, 2023, 1:15 p.m.
    ...the integrity of different targets and labels. You may want to check out a few different protocols to look...you’ve been looking for too!     References and resources References Bertiaux E, Balestra AC, Bournonville... two points within ~250 nm of each other. For reference, individual presynaptic terminals, which are jam...2020). Plus, it’s been demonstrated to work in different model systems, paired with SRM techniques, and...

  19. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ...known quantities of DNA. Additionally, an AAV reference, a virus with a known titer, can be used to confirm...both results would be valid. This is why the AAV reference is critical. It allows you to determine if your... PCR (ddPCR) does not require a standard or a reference, meaning saving on reagents (and time)! AAV titering...million.  After making the dilutions, they are transferred to a new plate containing the mastermix which...generator, you will have to be careful about transferring your droplets from the droplet generator to ...becoming a popular option for PCR experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and...

  20. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...Synthego has developed a new tool called ICE (Inference of CRISPR Edits) to solve this problem. This tool...efficiency and determine the profiles of all the different types of edits that are present and their relative...Contributions   The Contributions tab shows the inferred sequences present in your edited population and... an Indel plot (left panel) which displays the inferred distribution of indel sizes in the entire edited...average amount of signal that disagrees with the reference sequence derived from the control trace file. ...cut site, representing a high level of sequence difference. Traces The Traces tab shows the edited and ...

  21. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...colleagues reported that it is possible to reprogram differentiated cells into induced pluripotent stem cells (...original genome by integrating viral vectors are preferred. This is to avoid the chances of insertional mutagenesis...integration-free, karyotypically normal and are able to differentiate into various cell types of all three germ layers...https://www.linkedin.com/in/kmukherjeephd/.     References 1. El Hokayem, Jimmy, Holly N. Cukier, and Derek..., S., et al., LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-...Interview with Darrell Kotton Four Factors that Differentiate the Stem Cell Field Sign Up to Write for the...

  22. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    Published
    Oct. 19, 2017, 1:54 p.m.
    ...recombination between those sites. FRT sites and loxP sites differ at the nucleotide level but share an overall structure... Optimizing FLP FRT through smart engineering Different FLP versions – making FLP efficient The first ...efficiency to Cre (Kranz et al. 2010). These differences in recombinase efficiency are important to keep...inverted is flanked by two pairs of target sites that differ in their sequence. Recombination between the two...should be visualized at the same time from two different plasmids this approach becomes problematic as ...the genOway press release for more information. References 1. Atasoy, D., Y. Aponte, H. H. Su, and S. M....

  23. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...Genomic Tools) Program and I met scientists from 21 different labs who clearly also share this wonder of the...convergent evolution because electric organs arose in different branches of the evolutionary tree at least 6-7...cell line, then take the nucleus and put it in a fertilized egg to get edited embryos (replacing the nucleus...editing in the brain allows the squid to create a different protein. Did you know that most cephalopods die...behavior. For example, two similar species have very different social behavior. One is monogamous and one is ...new tools being developed and shared for many different organisms. Do you have a favorite organism you...

  24. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    Published
    Jan. 17, 2019, 1:38 p.m.
    ...critically important to be able to conveniently reference specific pieces of data stored externally in the...of research compiled in the ELN. Without such references the record of research is incomplete, making ...research Just as researchers need to access and reference external data sources, they also need to continue...data from the ELN directly into at least one and preferably more than one of these five general purpose repositories...spreadsheets, images, sequencing data, etc.) used and/or referenced in the experiment. Since most ELNs, unlike data...produced, collected, and analyzed. While the many different ELNs out there can help with searchability, record-keeping...

  25. How to Be an Excellent Trainee

    Type
    Blog Post
    Published
    Sept. 26, 2023, 1:15 p.m.
    ...mentor’: someone who provides career advice and offers direction on your scientific project. Your scientific...protocol you don’t understand the purpose of, or a different way of doing something then you’ve done previously...an active partner Everyone’s learning style is different, so let your mentor know what yours is! For instance...ask questions throughout training, while others prefer to review their notes, process what they’ve just... can also ask for a copy of the protocol if you prefer to take notes on a printed protocol, or maybe you... take training seriously, as reviewed above. Offer to help clean up and/or make necessary reagents ...

  26. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...) enzymes. Various other proteins included in different CGBE tools allow the researcher to direct the ...a translesion synthesis polymerase known to preferentially incorporate A bases across from abasic sites...recognize thymine (T) or cytosine (C). Several different teams succeeded with this approach around the ... gCBE is a deaminase-free C → G editor with a different editing window than existing CGBEs. This distinct...what tools the field will come up with next! References and Resources Glossary Here’s a quick refresher...respectively, to be replaced with another base. References Chen, L., Hong, M., Luan, C., Gao, H., Ru, G....

  27. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    ...CRISPR-associated transposition.     References and Resources References: Jiang Y, Chen B, Duan C, Sun B, ...concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the plasmids by...MUCICAT toolkit includes versions of plasmids with different induction systems, such as PT7, Prha, Ptet, and...integration frequency. (C) Increasing rounds of transfer on transposition-inducing agar lead to higher ...Capability to construct a strain library with different gene dosages to facilitate swift optimization ...

  28. PEI Calculator for Planning AAV Packaging Transfections

    Type
    Blog Post
    Published
    Jan. 23, 2018, 2:04 p.m.
    ...use the ratios below: 1 Transfer Plasmid : 1 pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap...Calculations are based on using 28 μg of total plasmid (Transfer, helper, and RepCap added together) per 15 cm ...the calculator in the comments section below. References 1. Choi, Vivian W., et al. "Production of Recombinant...Resources on Addgene.org Find AAV Packaging and Transfer Plasmids Here New to Making AAV? Check out Addgene...

  29. Streaking for Single Colonies: The Streak Plate Challenge

    Type
    Blog Post
    Published
    March 2, 2021, 2:15 p.m.
    ...He mixed together 10 E. coli strains producing different pigments, and tasked the students to separate ...over 20 years. Previously, the lab used three different species for the challenge, but one species always...Addgene is have the students transform them into different strains of E. coli,” says Westenberg. During the...transformation experiment, he plans to show how different genetic backgrounds of the host strains affect... Jennifer Tsang...

  30. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ...Gates are the parameters the machine uses to differentiate between variations in those factors. To understand...a little bit of theoretical background on the different parameters that your gating strategy will be based...measured: height (H), area (A), and width (W). The different measures are represented as FSC-A/SSC-A, FSC-H...cells, blood cells and more), each coming with a different size and structural complexity. After several ...still be gated out later. Based on personal preference, some people like to make stricter gates while..., unstained population, which I can clearly differentiate from the positive one (Figure 8). Note that ...

  31. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...mutagenesis. In addition, you may want to try different concentrations of template (e.g. 0.1-1.0 ng/μl...). Such non-complementary bases at the 3’-end interfere with plasmid reconstitution, and consequently ...relative to the original template). Due to the small difference in size, the fragments should be separated on... at extremely low frequency), and these could interfere with the function of the resulting construct. ...recognizes a unique site in the coding region (preferably near the 5’-end) and that also generates 4 bp... is a researcher at Cornell University.       References 1. Breslauer, Kenneth J., et al. "Predicting ...

  32. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    Published
    Nov. 25, 2015, 4 p.m.
    ...that, even though postdoc associations across different universities are trying to compile more comprehensive...path,” but individual institutions can have many different names for the positions commonly thought of as... life sciences name faculty-research as their preferred career outcome - see below). Stephan additionally...the training they receive as postdocs is not transferrable to other environments. Slide from Paula Stephan...slides can be found here). Importantly, the panel offered some great solutions to these problems and some...achieved. Of note, it was highlighted during the conference that just increasing funding for academic research...

  33. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    Published
    Oct. 2, 2018, 12:56 p.m.
    ...something you CAN choose and it will make a huge difference for your future career and life. You might see...amazing scientists, but the labs were profoundly different. My first rotation was with a mid-sized lab with...hone in on the factors that will make the most difference for your success. Gather data on the lab choices...careers?    Will you be encouraged to develop transferable skills by attending career events or being involved...your best performance and learning? Everyone’s different so give some thought to what atmosphere makes ...resources on the Addgene blog Check out our transferable skills guide Learn about careers in science ...

  34. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...their different migration speeds; after running the gel for some duration of time, these differently sized...time required for complete digestion varies for different enzymes. Many companies now sell fast digest enzymes...purification. In gel purification, you use a voltage difference across a gel matrix (usually agarose) to pull...sized pieces will be in different locations and can be cut out of the gel individually. There are a variety... plasmid DNA from the column using a particular buffer or water. See column manufacturers for more detail...

  35. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...and a screening approach, the Jennifer Doudna lab identified three different Cas12a inhibitors. Some of ...hisashii, BhCas12b that is functional at 37 °C but preferentially nicks the non-target DNA strand. The lab then... of stable genomic integration and is easily transferred via conjugation.   Read the paper in Nature... Jennifer Tsang...

  36. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    Published
    Dec. 29, 2020, 2:15 p.m.
    ...sharing even broader and began working with tech transfer offices to make reagents available to scientists... Proteins For Molecular Biology Experiments by Jennifer Tsang Chromoproteins are a subset of the fluorescent.... 7. 25 Work From Home Tips for Scientists by Jennifer Tsang These tips were written early on in the...Five DNA Quantification Methods to Consider by Jennifer Tsang The nanodrop vs. spec debate dominates ... Jennifer Tsang...

  37. A new platform for fast functional characterization of potentially cancerous mutations

    Type
    Blog Post
    Published
    May 7, 2019, 12:39 p.m.
    ...dependent cell lines and any mutations that that confer proliferation advantages will be revealed in the absence... a given mutation will have a different phenotypic effect in a different system. These plasmids can also...well-studied candidates. The possibilities are vast. References Ng, Patrick Kwok-Shing, et al. "Systematic functional...

  38. Addgene is Expanding Our Viral Vector Service!

    Type
    Blog Post
    Published
    Oct. 15, 2024, 1:15 p.m.
    ...use in this service.  At launch, we will offer five different serotypes for packaging: AAV1, AAV2, AAV5...the Addgene repository and their choice of five different serotypes.   Figure 1: The process of...that this ordering process will look slightly different from our standard process. We’ll walk you through... Preps will be available for request in three different quantities: 10x 100 µL (1 mL), 25x 100 µL (2.5...

  39. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    Published
    Nov. 27, 2019, 2:03 p.m.
    ...neuronal populations to consider. To tease apart how different neurons impact a certain output, neuroscientists... populations using viral strategies Combining different approaches can increase the specificity of cell...optogenetic payload in neurons that project to different regions of the brain. A more locally defined set...in their cell bodies. Optical stimulation in a different brain area can then reveal the cells bodies of...share experimental design and data. With these different approaches to open science, we aim to accelerate...

  40. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    Published
    Aug. 11, 2015, 2:30 p.m.
    ...NmCas9 and St1Cas9. Each ortholog was fused to a different fluorescent protein to create three colors. The...specificity of these orthologs is key: due to differences in the PAM sequences required by each ortholog...specific for telomeric sequences and showed that different fluorescently labeled dCas9 isoforms are efficiently...target sequence. They were also able to label two different pairs of chromosomes using gRNAs specific to sequences...Addgene are excited to see what’s coming next! References Multicolor CRISPR labeling of chromosomal loci...
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