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Showing: 441 - 460 of 1076 results

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

    Type
    Blog Post
    ... are equal. Created with BioRender.com. This kind of normalization is a nearly ubiquitous tool, but... assumptions that their loading controls are independent of these other sources of error. It’s important...housekeeping protein in your context. If you don’t find anything, you’ll need to conduct a control experiment...strongly influenced by unexpected variations in any individual protein. This ensures you are truly comparing...with β-actin (1:2000). Representative of three independent blots. B–C) Graphs showing the relative intensity...increasing concentration of β-actin in panel A. Indeed, the quantification in panel B is not especially...total protein are compared. While a higher slope indicates higher sensitivity, linearity is the most important...

  2. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...Expression Kit FREQ Seq PUF Assembly Kit Below you’ll find information about what these kits contain, how they...Biological Chemistry 290.8 (2015): 4728-4740. You can find more info on pCri in Mary Gearing’s blog post on...translation initiation sequence known as the Species-Independent Translational Sequence (SITS). This sequence ...composed of repeat segments that can be engineered to bind specific RNA bases. By swapping out these different...repeat segments, you can create a PUF protein that binds your RNA sequence of interest. The Zhao lab has ...contains all the variant PUF segments required to bind any 8 base RNA sequence. The segments are provided.... Using this system, you can create custom RNA binding proteins in as little as 3 days. Abil et al., 2014...

  3. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    ...to be binding at particular genomic sites when in fact they are just non-specific or indirectly interacting...is the go-to method for mapping where a protein binds genome-wide, and has been widely applied in many..., an enzyme-based protocol for identifying DNA binding patterns that overcomes some of the limitations...Protein A fused MNase is then added and Protein A binds Immunoglobulin G (IgG) thus targeting MNase to antibody...provides an excellent additional measure of DNA binding sites in combination with existing ChIP-seq data...with CAPTURE Additional Resources on Addgene.org Find simple lab protocols Browse CRISPR plasmids for ...

  4. Five Questions to Ask If You're Deciding Between a Master's or PhD Program

    Type
    Blog Post
    ...s-level job postings on a job search website like Indeed. Do the job descriptions sound interesting? Is ...work something you can see yourself enjoying?  To find master’s-level postings, try adding “master’s” to...filters to limit results to people that work in industries you’re interested in (i.e. biotechnology, pharmaceuticals...switch your field of work, both in academia and in industry. Advancing along a career path in these two fields...fields looks different however. In industry the career ladder is larger and there are more opportunities...Career advancement for PhDs, both in academia and industry, often means moving from the lab to a desk, although...on Addgene.org Check out Addgene's job openings Find eBooks from Addgene, including the Science Career...

  5. Embracing Serendipity: A Crucial Element in the PhD Journey

    Type
    Blog Post
    ... PhDs Making the Leap to Industry." He successfully transitioned to industry after his PhD and a couple...mental freedom. Practical approaches Engaging in mindful meditation, taking field trips, and spending time...gain fresh perspectives. Such activities foster a mindset that embraces the unexpected and encourages exploration...Serendipity is not just a stroke of luck; it's a mindset that can lead to unexpected discoveries and innovative...successful research career, whether within academia or industry. Albert Einstein once said: “There are only two...serendipity into your research but also cultivate a mindset that is adaptable, creative, and poised for groundbreaking...

  6. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    ...-4 colonies to ensure that you find the correct product. Once you find a colony with the appropriately...Posts Resources on Addgene.org Find Empty Vectors for Your Research Find Additional Protocols Browse Our... Some of these include, TA cloning, ligation independent cloning, TOPO cloning, one step cloning, and ...Li, et al., in 2011. This method is ligation independent, it does not require purification of insert or...simplified, purification-free, sequence-and ligation-independent PCR cloning method." BMC biotechnology 11.1 (...

  7. Management for Scientists: Giving Feedback

    Type
    Blog Post
    ...feedback when it is merited. Surprisingly, some people find this hard so get used to it by making it a concrete...However, I have also found that scientists seem to find it easier to give feedback in formal sessions using... Here are the 3 things that worked least well. Find out the Effect of the Feedback: How does this feedback...over, what would you do differently? Beware of indirect language You must be honest about performance ... mistakes I see is managers who hint at or use indirect language to talk about real performance issues...issues. Watch this little video to learn more about indirect language from Steven Pinker. As a manager you ...and clear. There is place in human discourse for indirect language, but it is not the way to go when giving...

  8. Fluorescent Biosensors for Measuring Autophagic Flux

    Type
    Blog Post
    ... autophagic activity, and are a more reliable indicator of autophagic activity. The aim of this post is...emits red and green fluorescence. Following the induction of bulk autophagy, Rosella is targeted to the ...interest to track the autophagy of that protein. Find the cytosol-targeted and mitochondria-targeted Rosella...nm to 440 nm serves as a measure of autophagy induction: when Keima is in the cytosol, this ratio is low... this biosensor can’t be used with fixed cells. Find the mitochondria-targeted Keima variant at Addgene... in yellow puncta but a decrease in red puncta. Find the mRFP-GFP-LC3 plasmid at Addgene! GFP-LC3-RFP-LC3...fluorescence suggests a decrease in autophagic flux.   Find the GFP-LC3-RFP-LC3∆G plasmid at Addgene! References...

  9. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    ...for scientists working at home, and you will still find this useful. Now that we’ve been doing this for ...safe space. No matter why, now is a great time to find a few colleagues to work together on career and ... “the ability to assess and initiate things independently” and “the power or opportunity to act or take...format is good for everyone including those that find events stressful. Video chatting makes it pretty...effectively is a valuable skill. There are many different kinds of grants that you can apply for. If you are in ..., your PI is likely working on a number of them. Find one to apply to or offer to help with lab grant ...Opportunities Open Education Database: 100 Places to Find Funding For Your Research University of Florida:...

  10. Summer SciComm: Science Storytelling

    Type
    Blog Post
    ...still invested in seeing how they do it.    Finding Conflict in Science  In all science communication...conflict into your work, but one of my favorite is to find it within the science itself - within that process...trying to understand your completely unexpected findings, or understanding why your experiment has not ...not worked…again…for the seventh time….   I find that once I sit down and talk to a scientist about their...paper was actually from a completely unexpected finding in a different experiment, this tool was developed...fodder for science communication. The trick is to find one or two roadblocks that sparked creativity, changed... a story around them. A good rule of thumb is to find the pieces that make you think, “Oh, that’s interesting...

  11. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    ...student hasn’t been asked the question: “Academia or industry?” Once academia was the clear answer with a well-defined...web-based, career-planning platform called the Individual Development Plan (IDP) and uses it to match qualified... qualified scientists to jobs in industry, academia, and government. This great resource allows the undecided...and other professions increases your chances of finding a well-suited job. EAs shouldn't be viewed as an...interests: Use career planning tools such as the Individual Development Plan to generate a list of potential... more information Follow people in academia, industry, and government on social media (Twitter, Facebook...potential job Join LinkedIn and other job sites to find open positions and view their required qualifications...

  12. Deep Dive: qPCR

    Type
    Blog Post
    ...single peak, indicating one amplicon and no primer dimers. (b) A shoulder peak indicating primer dimers...the right conditions and parameters, some people find it can detect even smaller differences, and it can...dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K. Filutowska; Selma...different types of pipettes and pipetting methods to find what’s most consistent in your hands.  When using...target in the reaction should be compared to an individually run sample when developing the protocol and ...and optimized until the multiplexed and individual values match across the board. If the samples or sample... e-book on applying MIQE guidelines! With these kinds of resources, you will soon become a qPCR master...

  13. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...targeting and sequencing millions of cells. For every individual sgRNA, you recover a minimum of 1,000 targeted... 5% off-target frequency is something to keep in mind, but likely will not have a significant impact on... many predictive online tools which can not only find gRNAs for you, but also score their probability ...probability of off-target events: CRISPOR, Cas-OFFinder, CCTop, and many more. While such software typically only...at mismatched guide sites, but not necessarily binding to these sites. Thus, these enzyme swaps will only...need to do your due diligence to ensure they are indeed a non-issue. Above we mentioned how to predict ...tested with some success. The major con is that binding by Cas does not always correlate with catalytic...

  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...expected fragments for that plasmid. This phenomenon indicates the presence of a multimer.   Fig. 1:...multimerization has been observed to occur in many different kinds of plasmids, taking many forms (Levene, 2009; Higgins... al., 2010; Cook et al., 2020). CGE uses laser-induced fluorescence (CGE-LIF) for detection and can provide...multimerization. One study by Bacolla et al. (2011) indicated that strain HB101 produced the highest yield of... at reducing multimerization and define recA-independent mechanisms. Avoid overgrowing the bacterial culture...-B DNA-forming sequences and WRN deficiency independently increase the frequency of base substitution ...26104344. Fishel RA, James AA, Kolodner R. recA-independent general genetic recombination of plasmids. Nature...

  15. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...seamless manner, leaving no sequences or mutations behind. Furthermore, piggyBac™ offers a large cargo-carrying...cells lines, including stem cells, without leaving behind any accessory DNA sequences [2], [3]. Simply put...integration [4], the donor cassette is removed leaving behind no accessory transposon sequences.  The use of ... plasmid after excision by PBx, thereby leaving behind only those cells with a Footprint-Free™ genome ...this method can be effective, a loxP site is left behind in the genome following donor removal, which alters...somewhere between one hundred and one thousand clones to find a cell line with the edit of interest. With the ...Cathomen, and K. Schwarz, “Zinc-finger nuclease-induced gene repair with oligodeoxynucleotides: wanted ...

  16. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    ... are interested in understanding the mechanisms behind Mycobacterium tuberculosis resistance to pyrazinamide... and recombination-mediated genetic engineering (find more information on bacterial engineering using ...Devarshi Gajjar, The M. S. University of Baroda, India  Devarshi Gajjar has 10 years experience in the ...expressing fluorescent proteins will be used to tag individual strains of bacteria. Their interactions and growth...University of Ibadan, Nigeriaunder supervision of Dr. Akindele Oluwatosin Adeyi, University of Ibadan, Nigeria...PhD Candidate working under the supervision of Akindele Oluwatosin Adeyi at the University of Ibadan. ...regulation and diabetes will be expressed from plasmids (find more information on bacterial expression systems...

  17. Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids

    Type
    Blog Post
    ...found yourself frustrated reading through a paper to find that there is insufficient information about which...also designed to be persistent and exist online indefinitely. PIDs historically referred to objects that ...identify a plasmid available through Addgene and find verified information about each plasmid item. Given...associated plasmid page on the Addgene website and find specific information about the plasmid: name, description...and more. The plasmid page also helps researchers find other publications that have used the same plasmid... Addgene ID # and the information necessary to indicate that this ID is managed by Addgene. How to cite...down to the bottom of this plasmid’s page you will find that the “How to cite this plasmid” section now ...

  18. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    ...represent the fraction of that total with the indicated function. One huge reason CRISPR has become such... left the arduous task of virus production to individual labs, but now we’re very excited to provide ready-to-use...an entire lentiviral production protocol only to find that the low titer of her prep makes it unuseable...target these widely studied signalling pathways. Find Validated gRNAs to use with your Cas9 Virus Pooled...uses nearly 80,000 gRNAs to target around 20,000 individual genes. We provide enough lentivirus for you to... lentivirus production protocol here and you can find more tips and protocols for working with lentivirus...Viral Service Page Check Out Our CRISPR Guide Pages Find Viral Vectors ...

  19. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    ...size of the insert. You can search NCBI for YGOI to find the particular reference sequence if necessary. ...The plasmid was digested with 2 unique enzymes (HindIII and BamHI) and run on an agarose gel. The resulting... (*). When the plasmid is digested with either HindIII and BamHI alone (lanes 4-5), there is a single ...size of the plasmid. The double digest with both HindIII and BamHI (lane 3) produces bands at 6kb and 1.2kb...bromide (EtBr) to your gel before pouring it. EtBr binds to the DNA and allows you to visualize the DNA under...vist Addgene's resource for plasmid verification to find additional tips and detailed protocols on topics...Resources: Addgene's Molecular Biology Reference Find More Protocols and Videos      ...

  20. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    ...novice and indicate why other neuroscience newbies might want to get into the field. Find Optogenetics...neuroscience but are afraid you don’t have the right kind of biological expertise to get into the field, there...require a bit of expertise and can be difficult for individual labs to make efficiently. For select viral vectors... to get some ideas for future experiments and to find tools that will make your transition into the field... If there’s a plasmid or AAV tool that you can’t find, you can always suggest a plasmid to us, request...Resources at Addgene.org Addgene's Viral Service Find Retrograde AAV Browse Optogenetics Plasmids ...
Showing: 441 - 460 of 1076 results