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We narrowed to 1,053 results for: FER

Showing: 481 - 510 of 1053 results

  1. How to Be an Excellent Trainee

    Type
    Blog Post
    ...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 ...

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

    Type
    Blog Post
    ...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....

  3. PEI Calculator for Planning AAV Packaging Transfections

    Type
    Blog Post
    ...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...

  4. Streaking for Single Colonies: The Streak Plate Challenge

    Type
    Blog Post
    ...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...

  5. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...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 ...

  6. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...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 ...

  7. Thoughts on the Future of Research 2015 Boston Symposium

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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 ...

  9. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...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...

  10. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    ...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...

  11. Addgene's Top Blog Posts from 2020

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  14. Addgene is Expanding Our Viral Vector Service!

    Type
    Blog Post
    ...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...

  15. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...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...

  16. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    ...the efficiency of Cre recombination varies for different constructs and cell types. Second, Cre may induce...from the standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding...enhance genome editing capabilities in Drosophila. References: Multiple new site-specific recombinases for ...for use in manipulating animal genomes. Nern A, Pfeiffer BD, Svoboda K, Rubin GM. Proc Natl Acad Sci USA...PubMed. Reduction of Cre recombinase toxicity in proliferating Drosophila cells by estrogen-dependent activity...

  17. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    ...Dr. Jennifer Doudna and Dr. Emmanuelle Charpentier. I was joined by Addgene Scientist Matt Ferenc (one...especially excited to attend this event because Jennifer did her graduate work one floor above mine in ...system Drs. Dounda and Charpentier met at a 2011 conference and decided to collaborate on what was a “curiosity... science to the non-scientists in the room. He offered the great analogy of comparing the CRISPR/Cas9 ...

  18. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    ...online on Addgene's CRISPR Library Reference page.    References: Genetic Screens in Human Cells Using...GeCKO library, each library has been enriched for different gene targets. These pooled libraries include enrichment...lentiviral libraries and their demonstrated application offer proof-of-principle that CRISPR-Cas9 can be used ...any sequence in the genome. Tips and tricks Wang offers this tip: “It's quite important, at every step ...

  19. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...at how variable Cas9-expression was across the different cells (Figure 3).   Figure 2: Cas9 expression...publication. Pro-Tip: 1. Use multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA...our website. There are a few reasons to target different regions of the gene with multiple gRNAs, but the...comparing the phenotypes of multiple gRNAs. Targeting different regions of the gene is also useful in case the...that would co-migrate on a gel) instead of two different sized digestion products (which could be easier...your way to making new scientific observations. References 1. Rago C, Vogelstein B, Bunz F. Genetic knockouts...

  20. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ... just find one that naturally occurs? This is different than the engineered approach used to make eSpCas9...SpCas9 or GeoCas9 RNPs were incubated in reaction buffer at temperatures ranging from 25-80 °C. Activity...incubated in human plasma diluted with reaction buffer for 8 hours at either 37 °C or 4 °C. Substrate ...gel. SpCas9 is stable when incubated in reaction buffer alone, but has limited activity when incubated ...are available from Addgene. Happy CRISPRing! References Harrington, Lucas, et al. "A thermostable Cas9...

  21. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...fundamentals. Our mission is to curate a one-stop reference guide for plasmids, so that you can spend less...their artificial nature, lab plasmids are commonly referred to as “vectors” or “constructs.” To insert a gene...Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the level of a chemical in a particular...plasmid use in the lab. Addgene’s Molecular Biology Reference includes information about molecular cloning, ...a plasmid vector, molecular biology tools and references, and how to maintain your plasmid stocks. The...

  22. Antibodies 101: Affinity Tags

    Type
    Blog Post
    ...antibodies, and more!   Resources and References   References Coligan, J. E., Dunn, B. M., Speicher, ...blot and immunofluorescence. GST  Glutathione S-Transferase (GST) is a 26 kDa tag which can be isolated with...Nonetheless, this tag can be purified with several different resin types and is a fairly small tag that is ...  Affinity tags vs. other tags There are many different types of tags, including epitope tags (FLAG, Myc...

  23. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ...tools for your next experiment! References and Resources References Wood, V., et al., Hidden in plain...an endogenous gene. Each trap uses a slightly different approach (described in detail below), but fundamentally...kidneys at a specific developmental stage you could infer that the gene which GFP was monitoring was being... these circumstances, trap methods may be the preferred screening option. Promoter traps Promoter trap...

  24. Cellosaurus: A Cell Line Information Database

    Type
    Blog Post
    ...specific compounds, links to publication references, and cross-references with cell line catalogs and collections...be adjusted based on query, reference, and shared allele search preferences to help you identify proximal...to learn more, check out the Cellosaurus FAQ! References Lorsch JR, Collins FS, Lippincott-Schwartz J....

  25. Running for Rare Disease, Running for FOP, Running for AJ

    Type
    Blog Post
    ... with a six-year-old boy named AJ Gonzales who suffers from Fibrodysplasia ossificans progressiva (AKA...first met AJ and his family, he did not seem much different than my second son of similar age. When he came...therapeutic antibody that binds Activin A and interferes with inappropriate activin-ACVR1 interactions...superpower for those running on behalf of rare disease sufferers like AJ. As the Blue Lantern Oath says: "In fearful...running on behalf of AJ Gonzales, a young boy suffering from FOP, raising money for NORD for the past ...

  26. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    ...antibodies for tagging purposes. Different antibodies bind different protein sequences called epitopes...infamous frankenstein, was constructed by grafting different parts together. By combining two optimized elements...living cells. To make these scaffolds specific to different targets, the scientists fused them to sequences...to build a more robust set of frankenbodies. References Ewert, S., Honegger, A., & Pluckthun, A. (2004...

  27. What Do I Do Now? Academic v. Non-Academic Career Decisions

    Type
    Blog Post
    ...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...of today’s postdoctoral training can directly interfere with a seamless jump into industry.” In his article...interact with scientists just like you What is different about academia? Publishing in peer reviewed journals...Constant worry about grants and renewals What is different about non-academia? Scientific publications are...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

  28. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...work demonstrates that different repair template strategies activate different HDR sub-pathways like synthesis-dependent...live updates! My most popular tweet from the conference is shown above. The blog post I mentioned, John...post-CRISPR cleavage When I first looked at the conference agenda, I was surprised to see that it would ...Having had time to digest my notes, since the conference, I think I may have finally grasped just how ...

  29. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    ...due to genetic drift and the use of genes from different microbial strains. It proved to be impractical... least 96% sequence identity when detecting a reference feature. For a coding sequence feature that may... could miss a common feature if the sequence differences exceed the threshold. This issue is addressed...often come in closely related versions that have different properties. To prevent misidentification, we augment...way for SnapGene users to apply features from a reference strain to newly sequenced strains through our ...

  30. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...cell-free transcription/translation systems (also referred to as TX-TL systems) have been gaining popularity...initial system by using a stronger promoter and different biochemical secondary energy sources to increase...XMAS-TREE. a) Cells are transfected with three different vectors and flow cytometry is used to sort populations...XMAS-TREE-based methods at various genomic loci in four different categories.     Brookhouser, N., et al. BMC...10.1186/s12915-020-00929-7    The CRISPR corner By Jennifer Tsang New CRISPR plasmids are always being added...
Showing: 481 - 510 of 1053 results