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Showing: 391 - 420 of 868 results
  1. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ... the library are differently sized, smaller plasmids may be amplified preferentially, skewing the composition...successful amplification? To provide a point of reference, Addgene commonly prepares NGS samples using both...overrepresented plasmids (plasmids that were amplified preferentially). You can also rank the plasmids by abundance...create a “cumulative wealth distribution” curve, or Lorenz curve. By calculating the area under the curve ...
  2. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    Published
    Aug. 28, 2014, 3:34 p.m.
    ...based on the predicted sizes and organization of different features within the plasmid. Restriction analysis...can search NCBI for YGOI to find the particular reference sequence if necessary. The example plasmid on ...fragment having increased intensity to serve as a reference band. The uncut DNA (lane 2) shows 3 possible ...in the gel well and the dyes provide a visual reference point so you can easily assess how far the gel...Eductional Resources: Addgene's Molecular Biology Reference Find More Protocols and Videos      ...
  3. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...far the T4 exonuclease proceeds: at the first occurrence of the chosen nucleotide, T4 will switch to polymerizing...efficiently repair the plasmid. Created with BioRender.com.   SLIC also opened up a new world of multicomponent...bacteria, which repair the plasmid. Created with BioRender.com. How does SLIC compare to other cloning ...method’s flexibility allows it to be used with different types of PCR-generated inserts. By harnessing ...updated by Emily P. Bentley in September 2024.  References and Resources Further reading Harnessing homologous...
  4. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    Published
    June 12, 2018, 11:40 a.m.
    ...chromosomal domains, or how stretches of DNA on different chromosomes interact with each other. Chromatin...Architecture and Chromosome Organization. These aren’t the only investigators in the chromatin field who... Note that histone genes frequently have many different variant names, not all of which can be listed ...Hutchinson JN, Sara SA, Ensminger AW, Fox AH, Chess A, Lawrence JB Mol Cell. 2009 Mar 27;33(6):717-26. Reporter-nanobody...me at the Keystone Chromatin meeting, especially Karen Reddy (@theReddyLab), Thoru Pederson, Joanna Wysocka...
  5. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    Published
    April 5, 2022, 1:20 p.m.
    ...commonly used in research laboratories to detect the different subclasses, but there are also secondary antibodies...secondary antibodies to detect multiple targets with different mAbs in the same experiment. The Trimmer Lab found...within the plasmid backbone to generate R-mAbs of different subclasses, including subclasses that were not... useful research materials and information. References and Resources Additional Resources on the Addgene...Addgene's recombinant antibody plasmid collection References Andrews NP, Boeckman JX, Manning CF, Nguyen JT...
  6. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    Published
    Feb. 1, 2022, 2:15 p.m.
    ...the wrong everything else.      References and Resources References Boston University Flow Core. Fluorochromes...standard curve, another quick experiment, or using a different vial and saving the poor, abandoned antibody for...assays involve multiple antibodies, each with a different signaling molecule, that all need to be detected...   Fig 2: Try to inversely pair fluorophore strength with protein abundance.    Second, you’ll ...
  7. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    Published
    Dec. 11, 2018, 12:46 p.m.
    ... scientific conference, whether you are attending in person or monitoring that conference via social media...American Society for Cell Biology (ASCB) 2018 Conference. I don’t exactly know how I was selected to participate...like Fast Company, opportunities to speak at conferences, and offers for postdoc positions. Check out ...s worth a read since many of the pointers are different from the other links shared in the post. And the...
  8. Communicating Your Science With Help From ComSciCon

    Type
    Blog Post
    Published
    July 19, 2018, 1:12 p.m.
    ...Sanders of ComSciCon, the Communicating Science Conference series for graduate students. I believe that...giving a public talk, ask a friend who works in a different field to listen to you practice.  Ask them what...organizations. At ComSciCon, our primary mission is to strengthen and support the network of graduate students ...addition to this annual event, graduate students in different regional and disciplinary communities gather at...Learn about careers in science communication The strength of story telling Science communication: 9 strategies...
  9. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    Published
    Nov. 24, 2020, 2:15 p.m.
    ...developers work in a streamlined manner. Now, in my current role at Oxford Nanopore Technologies, I work with... part of my job is getting to talk to so many different people! I communicate with customers, software...Often, product managers will have three or more concurrent projects in varying stages, which can get messy...meetings. Meet with my boss and coworkers, discuss current and future features being built for the analysis...blogger Rachel Rubinstein! Rachel Rubinstein is currently a Field-based Product Manager at Oxford Nanopore...
  10. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ... a screening strategy as well. In a screen, you aren’t killing off a portion of the cells like in a selection...You’ll want to pick enzymes that will give you different sized bands depending on whether the plasmid contains...using vector-specific primers, look for size differences. You can also use primers where one primer anneals...popular sequencing primers in our Molecular Biology Reference. Sometimes common  primers can't provide all of... Sanger sequencing Read the Molecular Biology Reference ...
  11. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...tools to further their research. However, tools aren’t very useful if you don’t know how to use them. ... ideas come from social media conversations, conferences, and questions our scientific support staff receives...line with Addgene style.        Fig. 2: Reference image (left) provided to our graphic designer ...for our blog posts, we usually make figures in BioRender or use appropriately licensed and cited figures... We are constantly reviewing the resources we currently offer to identify gaps and add to them as techniques...
  12. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    Published
    July 6, 2017, 1:59 p.m.
    ...organizers. My overall impression from this conference was that Cancer Researchers and Immunologists...Here I aim to compile a few highlights from the conference which reflect how cancer/immunology researchers...multi-pronged attack. Inflammation and tumorigenesis The conference opened with a very interesting presentation by...our Fluorescent Protein Guide. Throughout the conference aspirin and the use of COX inhibitors was discussed...about our free deposit and distribution service! References: 1. Pietrocola, Federico, et al. "Caloric restriction...
  13. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...Linearized Plasmids After PCR). Current site directed mutagenesis methods Current SDM methods rely on in vitro...those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested with methylated DNA-specific...common methods for site directed mutagenesis   References 1. Trehan, Ashutosh, et al. (2016). “REPLACR-... on Addgene.org Browse Our Molecular Biology Reference Pages Choose Your Perfect Plasmid Backbone Find...
  14. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...journal.pgen.1004086 Ghezraoui H, Piganeau M, Renouf B, Renaud J-B, Sallmyr A, Ruis B, Oh S, Tomkinson AE...repair are easily re-cleaved while indel products aren’t, so repeated cycles will favor accumulation of ...Comprehensive Cancer Center at UNC Chapel Hill.   References Bennardo N, Gunn A, Cheng A, Hasty P, Stark JM...2014) Is Non-Homologous End-Joining Really an Inherently Error-Prone Process? PLoS Genet 10:e1004086 ....
  15. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...protein expression journey! Resources and References References Nabet et al. The dTAG system for immediate...expression of essential proteins (when knockouts aren’t viable). siRNA or RNAi are small, double stranded...days. There’s also a CRISPR for that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 ..., and expression is tunable by titration with different amounts of tetracycline. Tetracycline regulation...
  16. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    Published
    March 3, 2020, 2:15 p.m.
    ...1013 - 1014 vg/kg, a dosage comparable to those currently used in human gene therapy trials. They further...reproducibility as scientists can assemble parts from different labs and experimental systems in their own research...mutants, determined protein expression levels from different promoters, and inhibited gene expression using...C-terminus of Cre (CreC) and the N-terminus of Cre (CreN) are fused to the truncated Phytochrome B (PhyBΔ...eye, muscle and epithelial cells, as well as in different stages of development, ensuring the breadth and...
  17. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    Published
    April 14, 2022, 1:15 p.m.
    ...reduced-in-size options often have reduced binding strength, half-lives, and/or specificity as well. They’...bispecific, meaning that each scFv can target a different antigen; binding of each can be either simultaneous...convert anyone to downsize their affinity reagents, trendy as that may sound. Validated antibodies are still... a scientist looking for that perfect fit!  References and Resources Additional resources on the Addgene...AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018). Drug targeting and ...
  18. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...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...
  19. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...Emily P. Bentley in October 2024. References and Resources References Demanèche, S., Bertolla, F., Buret...functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense... also causes outbreaks of food poisoning, the difference is virulence plasmids! This category includes...Mclellan, S. L., Willis, A. D., Comstock, L. E., & Eren, A. M. (2024). A cryptic plasmid is among the most...
  20. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...mammalian plasmid here and edit on! References and Resources References Kwart, D., Paquet, D., Teo, S. et...sequence constraints, or at least has looser or different requirements. Luckily, there are some available...necessarily more flexible than NGG; they’re often just different (e.g. NGAG, NGAN, etc.). Additionally, alternative...al). Additionally, SpG Cas9 with the same PAM preference of NG was isolated by the Kleinstiver Lab (read...
  21. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ...28032891. This is a different version of Split-BioID. BirA* is fragmented at a different site than in Schopp... This can artificially select against PPIs that aren't stable outside the native cell context. The Split-BioID...of the BirA* fusion matters! Schopp et al saw different levels of fusion protein expression and biotinylation...research? Plasmids are available from Addgene! References Schopp, I.M., Ramirez, C.C., Debeljak, J., Kreibich...
  22. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ...whenever you hear it (whether at a scientific conference or a karaoke bar), you probably want to sigh....methyltransferase 1 (DNMT1) to copy methylation marks from parental to daughter strands of DNA. When dCas9 is bound...nearly 100% demethylation without the confounds inherent to dCas9-TET, allowing one to study the causal... CRISPR-based tools for epigenetic editing.   References and resources:  Additional Resources on the Addgene...Addgene.org Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al. (2020) ...
  23. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ... can only carry around 4.5 kb. Different AAV serotypes have preferences for specific cell types. For example...end of an experiment series would behave very differently (distracted) from the ones tested at the beginning...Experiment Behavioral scoring systems (experimenter differences in behavior observation); experimenter blinding...viral service for their behavioral studies. References Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene...
  24. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    Published
    June 27, 2017, 1:03 p.m.
    ...or undetectable FRET, well-characterized FRET reference standards can be used to validate FRET measurements...sophisticated questions about cellular processes. Currently, FRET experiments can probe protein-protein interactions...Borghi 2016) to a selected protein. Overcoming current limitations to permit the use of multiple FRET ... diseases and even assist in drug discovery. References 1. Baird, Geoffrey S., et al. "Circular permutation...biomolecular structure and dynamics in live cells." Current opinion in structural biology 34 (2015): 52-59....
  25. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    Published
    Sept. 13, 2016, 2:30 p.m.
    ... to address the unique needs of students from different departments across the university and alterations...track postdocs nationwide. On this level, FoR is currently compiling a database of institutional responses...implementation of new strategies designed to cope with current increases. These two rulings along with progress...Howell -Founder of Howell Communications Philip Brenner, PhD - Assistant Professor of Sociology. University...public policy. Follow David on Twitter @driglar.   References: 1. Alberts, Bruce, et al. "Rescuing US biomedical...
  26. The Materials Science of Optogenetics Experiments

    Type
    Blog Post
    Published
    Sept. 17, 2015, 2:30 p.m.
    ...Several protocols have been written outlining the different steps and materials required for each [1, 2]. ...discussed above, each opsin variant has slightly different activation/inactivation kinetics. For example,...Biology from the University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne... or follow him on Twitter @derekpsimonphd.    References 1. Gutierrez DV, et al. Optogenetic control of...channelrhodopsin for activation of CNS neurons. Current protocols in neuroscience / editorial board, Jacqueline...
  27. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    Published
    Sept. 24, 2024, 1:15 p.m.
    ...Find the MagLOV plasmid here!   References and resources References Duret, G., Polali, S., Anderson,...just be an artifact and eliminate sources of interference, but the result persisted. (Even better, multiple...be developed from other fluorescent proteins.  Currently, the team is working to identify how MagLOV’s ... of many optogenetic tools. And, since MagLOV’s parent protein AsLOV2 has already been adapted into a ...
  28. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ...However, if your flow cytometer reads them out on different lasers, you should not need to compensate between... Guide Chart.  Pro tip! Some antibodies go by different names depending on your flow cytometer’s setup...look at the other channels you’re using first. A different fluorophore channel may be affecting your populations...first before you apply compensation values to your current channels. A notable exception If you have a sample...CD8+ T cells and cancer immunotherapies. She is currently a research technician in Arlene Sharpe's lab at...
  29. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    Published
    April 18, 2024, 1:15 p.m.
    ...previous Addgene CRISPR publications.  References and resources References Pyhtila, B., Kasowitz, S., Leeson...with CRISPR pooled libraries, although we used different categories (Figure 2C). Note that in Figure 2,...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...
  30. Of Myc and Men

    Type
    Blog Post
    Published
    Jan. 19, 2023, 2:15 p.m.
    ...antibodies, context is key.    References and Resources References Evan GI, Lewis GK, Ramsay G, Bishop...cell proliferation and tumorigenesis.  Using two different synthetic peptides corresponding to linear sequences... or the C-terminus, Evan, et al. isolated six different monoclonal antibodies (Evan, et al. 1985). Three...-9E10 variable regions and sticking them onto different scaffolds. But over the years, several alternative...
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