...people to take a break from their everyday tasks and make time for special projects? We borrowed an idea from...great success at accomplishing “we-should-really-make-time-for-that” type projects. Cross-team “Random...communication between labs. Scientists are often willing to make time for free food, and these lunches would provide...between labs, and people work hard at Addgene to make plasmid sharing as easy as possible. However, I ...work to share time with the people around us. It makes the quality of our work better and keeps us smiling...

... or set of plasmids. We hope that these articles make it easier for you to find and use the plasmids you...for in vivo expression, and/or produce vectors to make mutant cell lines or transgenic animals.   Wall...To expand the capabilities of Gibson assembly and make a more modular platform, the Jacks lab generated...cumbersome to use in a high throughput manner or make it difficult to quantify interaction strengths. ...Cas9-based AID tagging Seven years ago, Masato Kanemaki's lab successfully used the plant-based auxin-inducible...introduction of auxin range from 10-20 minutes, making AID an appealing tool for studying essential proteins...described in their recent Cell Reports paper, the Kanemaki lab overcame this hurdle using CRISPR technology...

...This is something I was expecting when I set out to make my first CRISPR/Cas9 gene edit. I wanted to inactivate...articles (gasp). To my delight, not only was I able to make the edits without any reagent-grade endangered Martian...Vinculin is a loading control. 3. In terms of making the monoclonal line: I had never done it before...important results (for example, if you needed to make a figure with the data). Using acrylamide instead...agarose to get sharper bands. Conclusion Overall, making a human cell line with a gene knockout was straightforward...who you are, you could quickly be on your way to making new scientific observations. References 1. Rago...

...smallest bacteria. The absence of a rigid cell wall makes them resistant to antibiotics and antibacterial ...interpretation. Also, the signals from the nuclear area make the detection of mycoplasmas difficult (Ligasová...contamination when you are working without gloves or masks. Make sure to change the gloves and masks regularly instead...line stock for mycoplasma contamination before use. Make sure there is no contamination present in the serum...disposable pipettes to avoid cross-contamination. Make sure you keep all the required equipment and reagents...Avoid long exposure of the cell culture to air and make sure to tighten the flask cap before placing them...

... an incredibly fast-moving field, and we want to make it easy for you to keep up with new developments...to update our web resources and plasmid pages to make them more useful and easy to navigate. We also updated.... Base Edit: Base editing plasmids allow you to make precise C:G->T:A or A:T->G:C edits in DNA without...Like RNA targeting systems, RNA editing does not make changes to the genome, thus providing an avenue ...CRISPR Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: ...

... and then condensing that and making it accessible to students.” Making science approachable is often ...eScience Labs.  “We work with faculty members to make sure the courses encompass what they want to teach...onsite, Caitlin can visit the operations team to make sure “that everything that’s going into the kit ...non-profit Fred Hutchinson Cancer Research Center. And make sure to subscribe to the Career Channel of the blog...

... science brick-by-brick”. Our mission is to help make science mainstream and to show people the very human...at these struggles will bring people together and make these daily challenges a little easier to overcome... As we started building, we thought – how can we make the connection between LEGOs and science more concrete...and sounds to synchronize with the images thereby making the videos more real. To give you an idea of the...amount of work, see how many pictures we take to make a walk cycle of one step. Challenges in the LEGO...

...this responsibility seriously. Every decision we make, every project we undertake and every service we... that our materials are affordable but enough to make us self-sustaining. With this unique model, we are...opposed to fundraising.   Aside from enabling us to make all decisions with the goal of  benefiting our scientific...ever curious what specifically drives the decision making process at Addgene, just look to our core values...curated, high-quality data and reagents Addgene makes it easy for scientists to deposit and request plasmids...

... programs and it seems to me that most Addgenies make an effort to avoid plastic bags and to conserve ...conservation in the lab One stingy science project was to make my own glass milk for purifying cut plasmid DNA ...amount of wasteful packaging.  Turns out, one could make a lifetime supply for about $10 US dollars worth...accessed blogs on BitesizeBio is the protocol for making your own Taq polymerase by Outreach Scientist, ...founder, Melina Fan, recently shared our protocol for making competent cells to save time and money. My other...

...Resource Identifiers (RRIDs) for each antibody, making it easy for you to determine whether any given ...been, a community resource that supports both toolmakers and tool users. We encourage you to share your...resource. We are excited to work with all of you to make this the best and most accessible recombinant antibody... are some ways that you can participate: If you make recombinant antibodies or other recombinant affinity...get started.  If you have suggestions about how to make the resource better or ideas for new materials to...

...DNA, but they still rely on the host machinery to make additional copies. ori sequences are generally high...replication machinery room to come in and get busy making copies.     Types of origins of replication ...plasmid type in your cells at a time? Do you want to make a lot of your plasmid? Is the gene toxic in high..., so if you need two plasmids for an experiment, make sure they have "compatible" ori's. See the table...more copies with the addition of chloramphenicol – make sure your growth conditions aren't working against...

...Gearing explains how Cas9 homolog, Cpf1, may make it easier to make gene edits at new genomic locations. 6.... Why and Where Do I Find One? If you need help making a transition into a new job or maybe to an entirely...gene delivery, but have limited packaging capacity making them difficult to use with CRISPR as the cargo....Your Genome Edit Once you’ve used CRISPR/Cas9 to make a change to the genome, you need to check whether...

...within the UV spectrum, while wavelengths over 700 nm make up the infrared (IR) spectrum. When photons are ...advantage of a brighter fluorophore? Strong brightness makes it easier to distinguish a signal from noise; therefore... together in the same color panel. In this case, make sure to compare the excitation/emission spectra ...www.bdbiosciences.com/en-us/resources/bd-spectrum-viewer.   To make life easier, always try to use fluorophores with...worry, there are strategies to resolve this issue. Make sure to check out our blog post on color compensation...and 2) BV421 is a very bright fluorophore, which makes it easier to detect the transgenic protein. We don't...

...the readability of your writing. It makes suggestions on how to make your writing less complex. While I...’s a list of tools and resources that might help make writing a little easier. Writing schedule tools ... should accept all of the suggestions Hemmngway makes (it wasn’t designed for technical writing), it’s...and you can share your library with others which makes it easier to work with collaborators. Zotero is ...

...well-understood standardized terms makes it a lot easier for organizations to make policy decisions on research...positive feedback on “our” MTAs, which sometimes makes people think our MTAs are different than other institutes.... Setting clear expectations from the beginning makes the sharing process smoother, more consistent, and...Stopwatch award and seek feedback as to how we can make our process easier for others.  If you’ve still ...

...action. Instead of rushing in, you take steps to make it more likely you’ll get your tasks done correctly...to employers because they save time and money and make employees more productive. Careers where you’ll ...applicants compared to the number of available roles). Make your resume stand out Before you begin to apply ... need to think of other aspects of yourself that make you stand out. For every role you need to show that...Grad Student to Entrepreneur Learn to Network and Make the Contacts for Your Dream Job Resources on Addgene.org...

...working toward a solution. At the end of the meeting, make sure to thank everyone for their participation and...through a meeting with no idea why you are there. Make the most of everyone’s time by considering the level...grows, so will the length of your group meetings. Make sure to keep them manageable. At Addgene, teams ...last meeting. Use some positive peer pressure to make sure everyone completes their assigned items to ...like to send around a Google doc where people can make comments. At meetings, we just focus on the parts...

...documentation helps make sure everyone is on the same page. Effective delegation allows you to make the best use...expected to DO all the work – she is expected to make sure the work gets done and done well. Involving...for failure by delegating is a bad idea. How do I make sure it goes well? As with most managerial responsibilities...mistakes will be made that could have been prevented.  Make sure everyone understands that no question is “dumb...

...goals. You must understand your own style before making this choice.  How big is the lab? While larger ... the position. Am I choosing the right mentor? Make sure your advisors will be good role models and ...non-academic career path?  If you are considering this, make sure to choose an advisor that supports both paths...is one case where the topic of your research can make your transition easier – choose a lab that works...always be in the zone, but even brief stays there can make all the difference. Choose a lab that brings on ...

...Gel issues usually come down to voltage and time. Make sure to load an appropriate protein ladder on your...issue and can’t figure out what’s causing it, try making fresh running buffer. Old or improperly made running...in our Technical Design of a Western Blot post to make sure you have the optimal gel for your experiment...transfer. If you are expecting low amounts of protein, make sure your reversible staining method of choice is... fairly quickly, even at 4 °C, so when in doubt, make up a fresh batch.  Antibodies We always recommend...the solution used to expose them. You’ll need to make sure the sensitivity of your exposure solution matches... amounts directly, stripping and reprobing. Just make sure to go from least to most abundant protein if...

...transgene has integrated and make genetic crosses with balancer chromosomes to make genetically stable fly ...needs. If YFG is thrillingly new, you might have to make a transgenic fly in order to study it. The process...to worry! In flies, phenotypic selection markers make it easy to spot flies with the appropriate transgene...important to note so that you know what to look for. Making your transgenic fly and downstream experiments ...

...the improvements we’ve implemented and how they’ll make it easier for you to find the plasmids you need....SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid maps far easier to ...identities of most features labeled directly on them making them much easier to quickly identify. Improved ... to paste directly into your notebook. This will make it easier for you to share plasmid information with...their functions. We hope that these updates will make it much easier to browse the repository and quickly...

...At Addgene, it is our mission to make it easy for you to share plasmids. To achieve this goal, we will...and perform standard quality control but will not make the plasmids available until you provide us with...by sequencing. Upload any support files that will make it easier for scientists to best utilize your plasmids...it appears in the Entrez Gene list, as this will make it easier for other scientists to find information...and Mammalian Expression). This information will make it easier for scientists to find your plasmid by...Please review all of the information on this page to make sure it is correct. You can easily update the information...

...support for your ideas, invite others into decision making processes, and even interview better! As a scientist...happened to me.) They happen to everyone! Everybody makes mistakes, encounters technical difficulties, and...deep breaths. Then, identify the specific things making you nervous–audience attention? fear of messing...messing up?–and use the three tips above to make a strategy that can help. Even something as simple as telling...learned this skill in its most difficult form, which makes it the perfect transferable skill. Almost every ...

...antibodies in a single experiment then you will need to make sure that you can easily distinguish each different...before you will not only have peace of mind as you make your purchase but also someone to turn to for advice...epitope target, source, and other information to make sure you’re not trying to choose between the same...different names.  If all else is equal, feel free to make the decision by price point, shipping time, or other...experimental considerations - no other factors can make up for an antibody that doesn't work for your application...

...gelsolin, but any nanobody sequence could be used to make a RANbody. RANbodies are expressed in 293T cells...cells due to their IgGκ signal peptide sequence, making them easy to harvest from cell culture media. Their...staining GFP, RFP, H2A2B   Are you ready to start making your own RANbodies? Great! All of the RANbody plasmids... RANbodies. If you know of a nanobody that would make a useful RANbody, consider leaving a link to its...immunodetection If you enjoy DIY lab projects, consider making you own DNA ladders with these plasmids from Penn...

... In this first example to the right (# this will make you sad),  someone has put in a request for a particular...hand, we can go back and optimize our procedure to make the whole process work more efficiently. Get on ... Enjoy your happy lab staff. Barcoded materials make everyone's life easier, the time and capital cost...case, the physical considerations of the laboratory make object composition a smarter choice than inheritance...anything other than Python or MATLAB. Choosing Python makes it easy to spin up a read-only Jupyter notebook ...

...verify the identity and quality of the plasmids we make available and provide some advice on the steps you... information in a binder or digital notebook and make sure you note the freezer location of every plasmid...typical reaction, or about 250 amino acids worth, so make sure the feature you want to verify is within this...perform Sanger sequencing from glycerol stocks, so make sure to select the appropriate service and read ... a large number of sequencing reactions, you can make your life much easier by selecting the right primer...

...process. Again, thanks to our partners at seqWell we make use of a pipeline that takes the raw data from each... is useful, but not 100% complete, we will still make this data available as a partial sequence with the... load it on to our new MiSeq and set up the run making sure to note the appropriate barcode for each sample...described by Joung et al. 2017, which we hope to make available as part of our OpenBio Repository sometime...K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller I, Mitsopoulos A, Fan M (2020...

...of this?” are all very valid questions, and often make an already complicated process even more difficult...program is the best fit for you and your goals. Making a List of Schools Creating a list of schools is... to highlight in the interview is another way to make the virtual environment more comfortable and help...presentation. It can help soothe your nerves and make sure you cover everything you need to during the...COVID-19, financial and logistical obstacles could make touring potential programs and universities stressful...

...the development of a simple and scalable means of making targeted genetic edits. Cas9 is a DNA binding protein...Traditional Cas9 regulators In addition to its utility in making targeted modifications to DNA, Cas9 can be reprogrammed...modifying DNA and of modulating gene expression makes it an invaluable tool in the interrogation of gene...potent Cas9-based transcriptional regulator without making any functional changes to Cas9. For instance, when...been found to retain some ability to induce Cas9 to make edits (AC, WLC, and JQ, unpublished results). An...

...improved learning outcomes and the opportunity to make new discoveries. Provost notes the importance of...excellent training in improvisation and the latter making him a more effective motivator (“be like Tony Robbins...thousands of undergraduates. The MCC develops and makes freely available technical and pedagogical resources...transform undergraduate education in the sciences by making a greater number of students active participants...Course-based undergraduate research experiences can make scientific research more inclusive. CBE Life Sciences...

...doi.org/10.1038/s41586-019-1323-z Peters JE, Makarova KS, Shmakov S, Koonin EV (2017) Recruitment of CRISPR-Cas...array. Workflow and technical considerations Making insertions in bacterial genomes with the V. cholera...Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, Zhang F (2019) RNA-guided DNA insertion...doi.org/10.1126/science.aax9181 Strecker J, Ladha A, Makarova KS, Koonin EV, Zhang F (2020) Response to Comment...

... high-speed internet. Before you begin any work, make sure you know if your audience will be able to access...if needed. And since we live in the digital age, make sure to research any software you may need: cost...communications you create. Wildlife biology and ecology can make for beautifully compelling photos and videos, while...things that cannot be seen with the naked eye or to make things more accessible by including, highlighting...Well-developed content creation platforms.  Algorithms can make it difficult to reach people who are not already...

...updated in several years) or they make significant changes that make it incompatible with our software...a problem, but it is these very differences that make our software so great. If we all came from similar...a bag of flour is not the only thing you need to make a cake. The flavor, shape, and texture of a cake...understanding the hows and whys of our users, we are able to make informed software design choices, resulting in software...

...structure and function of Cas13a with a focus on what makes this molecule unique, and the various applications...nuclease Cas13a was originally identified in 2015 (Shmakov et al., 2015). They were using Cas1, a gene commonly... in length and does not require a specific PFS, making it a very flexible cleavage system. LwaCas13a is...excited to see this group of enzymes continue to make a splash in the CRISPR field. We feel that the culture... Find plasmids from this paper at Addgene. 6. Shmakov, Sergey, et al. "Discovery and functional characterization...

...regulate the Z pathway?" A well-defined outcome makes it easier for the search committee to understand... to be overly ambitious – a common mistake is to make each aim into a 10-year research program. It's a...communicate your story with only the most salient details. Make the most of your limited speaking time. Beyond ...vital task of engaging your audience. You need to make your audience care, to want to know the answer to...Creating a coherent unified story and model can help make your ideas easier to understand and more exciting...

...organism of choice. Unfortunately, codon preferences make it so you cannot choose among the possible codons...therefore stall at inappropriate locations or fail to make it through the entire transcript resulting in the...properly. You might then think, “Okay, I’ll just make sure the abundances of the codons I choose for the...surrounding codon choice, you should have some method to make sure the proteins you produce function properly....choice, you should perform a functional assay to make sure your protein isn’t forming inclusion bodies...

...targeted by specific sgRNAs. Its RNA-guided nature makes CRISPRa much simpler to implement in genome-wide...the SPH activator paired with AAV sgRNA delivery make the SPH mouse an attractive tool to probe gene-function...screening strategies Although the tools described above make CRISPRa more accessible to neuroscientists, not ... in vivo applications, the tools described above make CRISPRa in the brain relatively easy and accessible...overexpressing genes within discrete cell populations makes Cre-dependent SPH mice an attractive alternative...

...clear plans to connect Genotype to Phenotype thus making progress on the NSF “G to P grand challenge.” Creating... are great for studying parenting strategies and make important ecological indicators since species live...possible to actually silence this gene with RNAi to make a polygamous species monogamous. Anolis sagrei (...CRISPR/Cas in lizards. The Menke Lab was able to make an albino lizard with targeted mutations. Now what...this to edited those brown marmorated stink bugs to make them pink. We’re excited to see new tools being ...

...exclusive access to data and enjoy reasonable time to make sense of it, with their decision to publish setting...the clock for others to dive in. The real cost of making data accessible A more reasonable concern with ...raised by researchers worried about the effort to make the underlying data easily available, organized ... the scientific community is thus not averse to making data available, but there remains concern about...reader as possible. Rather than adding to demands, making effective data sharing the norm should open up ...

...through some of the most important considerations to make when planning and performing an IF experiment. ...antibodies in a few different species and colors makes it easy to perform multiplexed IF. By using multiple...Some of the most important considerations are to make sure your secondary 1) will recognize the species...Database and CiteAb. The antibody you choose can make or break your experiment, so take advantage of all...your experience, positive or negative, will help make antibodies more effective and reliable in the future...

...Two plasmids that can be used to make inexpensive 100 bp or 1 kb DNA molecular weight ladders were recently... plasmid prep protocol, it cost less than $10 to make 1000 lanes worth of the 100 bp or the 1 kb ladder... than commercial ladders. Are you interested in making your own DNA ladder? You can find the Tan Lab’s...Protocol Learn How to Analyze Your Restriction Digests Make Your Agarose Gel Other Resources Read about Gel...

...calculator can do the math for you and it’s simple to make adjustments for the type of tissue culture dish ...dish and number of dishes used. Here’s to making AAV packaging a little easier! Click here to download the... spreadsheet: We hope that this calculator will make it easier to plan your AAV packaging experiments...AAV Packaging and Transfer Plasmids Here New to Making AAV? Check out Addgene’s Step-By-Step Protocol ...

...available the potential mentor is. If the person makes time for you, as needed, this is a strong indicator...time and resources. You typically have 4-5 years to make discoveries and publish papers to position yourself...most exciting story I can tell with my data?  Can I make better slides that more effectively tell a story...present your research. Face to face meetings can make favorable impressions on potential manuscript or...minimal processing, and low in empty calories will make you feel better and help maintain your health. ...

...twenty graduate student volunteers worked hard to make this Saturday event a success, a true testament ...communication gap between scientists and the public and help make science more transparent. Second, SITN helps graduate...intelligible scientific resources. SITN hopes to make DayCon a yearly event and to expand the conference...is a great idea and a group of people willing to make that idea a reality! How do you share your science...

...Tools for Making Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for making scientific...Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection by Joachim Goedhart Guest..., and fusion location, there are many choices to make. 2. Prime Editing: Adding Precision and Flexibility...

..., I’m picking two posts that describe methods to make single base edits in DNA. The first, contributed...a thorough walkthrough of how you can use PCR to make small sequence changes to plasmid DNA. This an oldie...CRISPR endonuclease (dCas9) fused to a deaminase to make the transitions at target sequences specified by...informative post. Interestingly, this technology may make some uses of Site Directed Mutagenesis by PCR obsolete...

... a job you wish you knew more about – how do you make a connection? An Informational Interview is a great...relationship building through Informational Interviews. Making Your informational interviews productive Informational...phone or video call. If you do decide to use Skype, make sure to follow the rules of successful, remote conversations...many good lists of questions on the internet. To make the most of the opportunity, use a written list ...

...Tip 1: Use color coding, labels, and post-its to make finding your most used protocols & references easy...for you) and place them on an easy to reach shelf. Make sure all notebooks are visibly labeled on the spine... tabs (for different experiments or projects) to make it easier to know where to start your search. Then...track the revision history. Now you’ll be able to make updates to protocols in real time. You may also ...