... digest tips and tricks: The following tips will make it easier for you to obtain a useful and informative... recognition sequence. Most enzyme manufacturers make High Fidelity versions of the endonucleases and/...before loading them. The glycerol in the buffer will make sure your sample settles in the gel well and the...wrong! Quantify your DNA. Loading too much DNA will make it difficult to obtain crisp bands and analyze the...

...efficiency. Check out Addgene’s blog to learn about making your own competent cells and our protocols page... encodes a green fluorescent protein gene which makes the bacterium fluoresce green. Comic by Maya Kostman...Lytic phages hijack the bacterial hosts machinery to make more viral particles. Eventually the cell lyses ...bacterial chromosome and allows the donor bacterium to make a small “bridge” or sex pilus that attaches to the...

...sometimes called “VUS”) function is unknown. How can we make a decision for each patient if the majority of information...high-throughput manner and therefore push us closer to making cancer precision medicine a reality. Challenges...have worked closely with our Addgene colleagues to make these available to the scientific community. Functional... Institute of Harvard and MIT and is focused on making precision functional genomics a reality for future...

...Zetsche, B., Gootenberg, J., Abudayyeh, O., Slaymaker, I., Makarova, K., Essletzbichler, P.,  Zhang, F. (2015... diagram below for an example of how this works. Make sure to order 5’ phosphorylated oligos or treat ...single plasmids are generally more efficient. Pro-Tip Make sure to reverse the orientation of the crRNA’s direct...

... explore small-scale opportunities where I could make mistakes and learn from them. I started at my previous...Beyond Academia: Stories and Strategies for PhDs Making the Leap to Industry." He successfully transitioned...Resources  Resources for aspiring managers Books: The Making of a Manager by Julie Zhuo Radical Candor by Kim...Managing vs. Leading Management for Scientists: What Makes a Good Leader Anyways?  Transferable Skills Guide...

...think. So, if you’ve ever wondered what goes into ‘making virus’ or what gag and pol stand for, we demystify...recombinant adenovirus is missing the endogenous E1 gene, making it replication deficient. This is typically supplied... some of the plasmids in our collection. Want to make a vector construct on your own? No problem, we are...discussed are compiled for reference below. Happy virus making!  References and Resources Kulanayake, S., Tikoo...

...value. Types of biosensors So now that we know what makes up a biosensor, we can discuss what flavors of sensors...difficult to achieve in whole organisms, but AAVs make it possible. In AAV systems, different serotypes...expression and downstream monitoring. Nervous about making a virus for your biosensor? Don’t be! We have you...viral resources and safety tips. Still worried? We make virus with some of our most popular biosensors already...

...feature, you can compare different Cas enzymes to make sure you are getting the desired features and cutting...and discover orthologs with similar properties (Makarova et al). CasPEDIA is an evolving database, and... to it! The CasPEDIA Consortium will, of course, make efforts to keep it updated with new entries and ...Structural Biology. https://doi.org/10.32942/X2C31F     Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019)...

...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...Parton Lab recently created a flexible system that makes use of pre-existing transgenic lines, termed BLITZ...system provides researchers better flexibility and makes the approach more accessible, as it reduces the ... be packaged into an AAV system. dCas9-SSAP will make the editing of kilobase-scaled sequences more efficient...

...character you painted in your personal statement. Make certain to choose your recommenders based on your...individuals are inundated with requests. The easier you make it for them, the likelier you will end up with a...lighting for a smooth performance. Most importantly, make certain that everyone you live with is aware of ...consistent with your personal statement, while you make the ultimate case: why you? 7. Strength in numbers...

...variety of educational resources in an effort to make scientific knowledge broadly available.  The ability... return the matching resources in an order that makes sense to the user. The devil is in the details of... of course: what does “best” mean? and what “makes sense” to a user? Moreover, there are many ways a user...institutions. There are many more technical details that make our new search engine unique. Most importantly, ...

...will be a positive asset throughout your career? 1. Make scicomm practice a priority The good news for young...of the work you presented may also be unclear. 2. Make the audience your primary focus When writing a paper...address that 'why' question effectively, you need to make your audience the focus of every sentence you write...know this? How will they react to it - what will it make them think? Does this lead to the conclusions I ...

... task will keep you from repeating your work or making similar mistakes in the future. Stay organized...camelCaseFormat since spaces between each word can make it difficult to accurately call a file. Version...your exact changes. Alternatively, you can simply make a note in your methods section about the edit that... Your Experiments Try out the CRISPR Software Matchmaker Additional Resources on Addgene.org Find Protocols...

...behind. While commercial kits are available for making small point mutations, large deletions/insertions...Current SDM methods rely on in vitro assembly. To make a point mutation, you design two oligonucleotide...instead of relying on in vitro assembly, they could make the E. coli do the work by using the principles ...-12 kb range. REPLACR-mutagenesis also requires making your own electrocompetent cells in which the phage...

...between blue and white colonies. Take care in making your plates: X-gal is light and temperature sensitive...autoclaving. If spread on top of pre-made plates, make sure it is evenly distributed and allow sufficient...specific analysis, like PCR or restriction digest. Make sure you use a proper E. coli strain (i.e. contains...JM109, STBL4, JM110, and Top10 are a few examples. Make sure you use a proper plasmid (i.e. contains the...

...how the new kit works, the research the kit now makes possible, and how it compares to his CRISPR/Cas9...step. The kit also has other unique aspects that make it especially well-suited for use in stem cells....hands of pretty much all investigators. Our kit makes it extremely simple to design TALENs. It only takes...apples-to-apples comparison of all those mutations. If you make induced pluripotent stem (iPS) cells from patients...

...experiments, and here we will focus on a unique gene that makes the popular GatewayTM method possible: ccdB. But... more about it? Read on to find out how ccdB can make your cloning experiments a little easier. One of...contain your insert of interest. Simply stated, ccdB makes cloning easier by selecting against vectors that...not contain the recombinant plasmid and therefore makes the cloning process much more efficient, as one ...

...scientists commonly rely on RNA isolation kits to make their life easier. Recently, we published a blog...cold and work quickly to mitigate RNA degradation. Make sure to use DEPC-treated or RNAse free water. If..., 2006).  View our protocol for RNA extraction! Making the simple protocol even more foolproof (still ... if you need really clean RNA, it’s important to make sure that when extracting, to only take the aqueous...

...properties of the fluorophores you are considering to make sure that they’ll be compatible with your instruments...properties allow for efficient signal amplification, making biotin conjugated antibodies good options for detecting...antibody. 2) Endogenous cysteines, such as those that make up the disulfide bonds between antibody chains. ...antibodies yourself, there are kits available to make the process quick and easy. Before getting started...

... see who you are when you look at their profile. Make your professional contact information available.... be important for a good friend, but if you are making a professional inquiry, you always want to read...type of connection can help you gather information, make introductions, and give advice for your interviews...should be as good or better than your resume Don’t make the mistake of having a shoddy profile and not updating...