...NB) were generated in pMAK:  pMAK461(NBαRabitb-IgG-Fc, Addgene #140699) pMAK462 (NBαMouse/Rat-IgG1-Fc...#140700) pMAK463 (NBαMouse-IgK, Addgene #140701) pMAK464 (NBαEGFP, Addgene #140702) The pMAK461-64 plasmids...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...which alleviates the inclusion body problem and makes nanobody production in E.coli as simple as following...miniprep protocol!  First, the authors created the pMAK backbone by modifying pMAL-T-Avi-His/BirA, a vector...used for in vivo biotinylation via BirA and AviTAG. pMAK contains the signal peptide of MBP, followed by ...

...receive such a call, make sure to get that person’s contact information and again, make a personal connection... the hundreds of others that did the same thing? Make an effort to meet people. Attend smaller talks within... CV or resume). If you receive a card in return, make sure to follow up with a professional “thank you...company. It does, however, offer you an opportunity to make a personal connection; the importance of which will...there, so that you can tailor your CV accordingly. Make it stand out with key words and by putting your ...’ll find a variety of listings. Post your CV and make it visible, which will sometimes result in a recruiter... a traditional training program, and you hope to make a career in academia, you are likely in good shape...

...goals. These tools remind us of our daily tasks, make our days more effective, and our schedules more ...research. Electronic lab notebooks (ELNs) are here to make lab work much more reproducible, convenient and ...required every once in a while, so it is good to make sure you’ll be able to do it when needed. How many.... If these are important in your lab, you should make sure your ELN allows you to assign specific permissions...ownership should be yours (your institution’s), so make sure you don’t use ELN providers that take any ownership...set for every member of your team. Do you need to make sure your research can be patented? I.e. is automated...difficult to figure out all the ways a given ELN can make your lab life easier. If you can talk to an ELN ...

...Coaching can be an excellent tactic for scientists making a career shift or who wish to improve their current... committed to. Finally, hiring a paid coach will make you take the experience more seriously and I have...good ones take this responsibility seriously. As I make my rounds speaking at research institutions and ... Sometimes only a few hours of coaching time can make an enormous difference so don’t assume that this... with. Talk to the coach about their “style” and make sure this is a style that will help you. A good ...good coach usually will be a clear communicator and make direct suggestions for improvement and change. ...launched an online career coaching program that might make her coaching more affordable for you. There is also...

... as well as the instructions on how to make your own, will make it easier for you to use genomic tagging...alleninstitute.org. A classic challenge in cell biology is making sure that what we observe through the microscope...and result in a ubiquitous background signal that makes it hard to visualize the precise location of the...cellular structures, we provide guidance on how to make your own gene-tagging constructs below. Read on ...process following the double strand break CRISPR/Cas9 makes at the target site. The whole segment—a GFP tag ...cells. As a precautionary measure, it’s also good to make a few single base pair mutations in crRNA target...and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing the tag...

...nice if you can afford them). I am talking about making sure the audience is interested, educated and talking...with room for fortuitous improvisation. You want to make it look effortless to the audience, but the organizers...moderator should have a well-planned approach to making the event interesting and engaging. Long before...funny, provocative or interesting stories that will make for a fun discussion. People are much better at ...interesting questions and topics in advance is key. Make sure all your panelists have the correct date, location...as it approaches. Room set-up and logistics To make a really lively discussion, I strongly suggest asking...colleagues to events for this purpose. If possible, make sure the panelists are prepared to stay around to...

... in the world knows it—to be the first person to make a discovery. I can’t tell anyone else what the “...cases, don’t enjoy, is worthwhile and can help us make decisions about what we want to do in the future...only demonstrates initiative and drive, but also makes for an experience more tailored to personal interests...professors about their research, and learn about what makes them excited to do what they do. I would also highly...eggs in one basket. There are many factors that can make or break a lab experience. Previous Addgene blogs...want to dedicate your time and energy to. How can I make the most of my experience in the lab? Ask your questions—especially... I’ve been afraid to ask “dumb questions” and to make mistakes. Ultimately, though, scientists are all...

...produced in the lab. The first decision you must make is which system you will use, or more specifically...specifically, the type of cells that will be used to make the virus. Viruses can be produced in many different... by the specific type of virus you are trying to make. For example, bacteriophages will need to be produced...methods, understanding the differences can help you make an informed decision. Mammalian cells Mammalian ... they contain some of the necessary machinery to make these viruses. The cells are also easy to cultivate...mammalian cells. However, it is much more common to make lentiviruses and AAV in-house. High-titer adenoviruses... that specifically infect insects. This is what makes insect cells a popular choice for protein production...

...lead to hurt feelings, low morale, poor decision making, and stalled projects. You want to identify and...experiments to address a reviewer’s concerns, you are making your hypothesis stronger. When you share data at...re having--that conflict you are resolving--will make you a better scientist. And some amount of conflict...miscommunications in the future. Assumptions: Making incorrect assumptions can be a direct cause or ...or consequence of lapsed communication. Making some assumptions about how something should be communicated...know that a person or team should be involved in making a decision and you see no record of their input...in question to a specific person or team. What Makes You a Good Conflict Resolver? The first and foremost...

...information about AAV, read our AAV guide . Can I make a stable cell line with adenovirus? No, adenoviral...

...much for how bright you can make a reaction but for how sensitive you can make a reaction. NLuc requires...proteins to make better reporters and the second couples NLuc to fluorescent proteins to make better biosensors...interesting construct you wished to use, you could either make it yourself or you could “clone by phone”. One of...tactics for getting the plasmids he wanted. Addgene makes it so much easier to share your constructs from ... however NLuc’s emission peak is at 460 nm thus making it non-optimal for in vivo imaging. The solution...the fluorescent donor for a bioluminescent donor, making BRET probes. Here the bioluminescent donor is unaffected... fluorescent proteins has allowed researchers to make innovative tools not possible with other luciferases...

...genome is 1 in every 256 base-pairs of DNA sequence, making it very useful for genetic engineering approaches... and gene editing. How does the piggyBac™ system make it easier to generate transgenic cell lines? One...transgenic cell lines. The traditional method of making stable cell lines is to transfect, select, and ...negative impacts on fitness. One consideration when making a transgenic cell line with the piggyBac™ system...However, several challenges still remain, such as making gene edits completely seamless (i.e., so no other...piggyBac™ transposase (PBx), which contains mutations making it competent for excision, but defective for integration...larger piggyBac™ donor vector, it is possible to make the ITR and selection cassette disrupt the target...

... As a member of the Quality Control team, I help make sure the plasmids that go out our doors are correct...assistant in a lab at the university that I went to. Making that switch from being a grad student to an employee...she didn’t want to take it. “I realized if I don’t make it to med school it’s not the end of my life, even...programs. I thought, ‘Let me pursue this, let me make a slight change so I can have flexibility and be...is incredibly helpful to me to know that you can make big changes in your career path and still find comfort...never work a day in her life. She said, “it now makes sense because I love working in the lab and being...people and for the people, feeding the people, and making them get outside of their comfort zones. I came...

... to read it, give it back to you, and for you to make the relevant alterations before submitting - handing...otherwise why would you be interested in it?) so make sure that comes across. Check out some in-depth ...agencies want value for their money so you need to make it clear that you will be doing your research in...routinely isn’t common to non-scientists and may make your perfectly reasonable paragraph unintelligible... about what I hope to discover. This section may make the difference in whether you get funding or not...Improve Your Science Communication Skills 5. Practice makes perfect, apply for small grants too There are actually... awarded money to carry out research, be sure to make it clear what the outcome of the work was. Ideally...

...to Ireland and take a chance as entrepreneurs. To make things more interesting, we are working in a new.... Being unable to publish or attend conferences makes it very challenging to compete in today’s tough ... a lot of time talking about what we could do to make ourselves marketable to the wider world and occasionally...Keira’s an optimist, I’m a realist, and together we make a pretty good team. We have confidence that we can...the lines of “I hate GMOs, but what you are doing makes sense. I can support that.” That was interesting...technology we will need, and the business training to make the most of it. It has also allowed us to really... we enjoy and see a future in. And we decided to make education and outreach a cornerstone of our business...

...this make things more manageable, but it also keeps stock solutions from getting contaminated. Make sure...minutes before you start any tissue culture work and make sure that clean air is flowing in. The vent for ...not be used as storage cabinets. Clutter not only makes it difficult to clean your workspace properly, but...disrupt the laminar air flow around your work area. Make sure to only bring required materials into the hood...stay out too long While you’re preparing things, make sure your cells don’t stay out too long. Cells usually...indicates that the cells have grown enough, are making their surrounding media acidic through the secretion...all but one in liquid nitrogen. The frozen vials make up your master stock, while you can use the remaining...

...more easily identify what to delegate and to whom (make sure that person has time for it!). Then schedule...not a clear deadline. Finally check-in regularly, make yourself available, and encourage questions. Unless...opportunities. I still need to check myself sometimes to make sure I am delegating well, but it does get easier...grateful for the feedback, but at least you can start making changes for the best. Positive feedback is equally...experienced to guide you - is valuable. Protect your team Make you team feel safe. Your team members need to trust...to encourage them to speak their minds. They will make mistakes and you will take responsibility (within... and help you grow in your role. The latter will make you feel discouraged and not valued which will undoubtedly...

...you use depends on your preferences. Either way, make sure to test your colony PCR primers before using...insert or if the insert is in your plasmid Have to make a new primer pair for each insert Backbone-Specific...plasmid Great for blunt end cloning You have to make a new primer pair for each insert since primers ...positive ones. Regardless of which method you choose, make sure to use the appropriate antibiotic for selection...polymerase is sufficient. Controls: Controls can make or break an experiment. The best controls for a ...products on an agarose gel to determine their size. Make sure to use an appropriate molecular weight standard...doesn’t mean there aren’t mutations in your insert. Make sure to submit multiple positive clones for sequencing...

...manager quickly make the connection between your skills and the needs of the job. Make it clear that you...successful, reproducible assay. That information can make you a much stronger candidate!  Additional uses ...cover letter to explain why your transferable skills make you a good candidate for the position. You can also...could talk about how your experience with qPCR would make it easy to master ddPCR.  Finally, it’s a good idea...highlight what specific skills, qualities, or interests make you a good candidate for the job. The body of the...should be one to three paragraphs. This is where you make the case for why you are a good candidate, explain...start from a standard version, but take some time to make it fit the job you’re applying for. The cover letter...

...philosophy Customer service can be a hassle. You finally make it through a 20-step phone tree, only to be left... customer support with self service We strive to make our support team available whenever you need them...not have the chance to stop what you’re doing and make a phone call or write out an email. You need help...resources, we will continually update the Help Center to make sure that, whatever your support needs are, we’ve...ll navigate to articles answering your questions making it easier than ever to get the support you need... regular site, www.addgene.org. We’re excited to make these new features available to the Addgene community...valuable feedback. This is Addgene after all, and making your research easier always comes first. Resources...