...viruses, so they are resistant to alcohol-based disinfectants. A 10% bleach solution should be used to clean...from workspaces. Speed: AAV transduction experiments are relatively quick. We have found that expression...benefits make AAVs useful for a wide variety of experiments, care still needs to be taken when selecting ...complexes are so robustly expressed that anatomical assessments can be easily performed too. While I implied ...transport of AAVs is a potential problem in many experiments, a group of researchers has identified this feature...used to treat disease in humans and additional improvements are being made to make them even more useful...Made in Heaven: CRISPR/Cas9 and AAV Viral Vector Elements Additional Resources on Addgene.org Find AAV...

...new NEB kit includes a step that both fragments the DNA and prevents it from forming concatemers later. ...services and expertise without breaking their bank accounts. When I heard about how LabCentral aims to enable...must first fragment a DNA sample into many small components. Without proper processing, these small DNA strands...produced high yields of quality libraries from small amounts of DNA i.e 100 pg. At the start of the workshop...itself. This was great because it forced workshop participants from different biotech companies to interact...

...smaller fragments that are then barcoded for identification. This mixture of barcoded fragments is known...help unlock the potential of modern sequencing instruments by enhancing the efficiency and simplicity of...known as a *library*. Parallel sequencing of the fragments within a library allows researchers to reconstruct...together to create a single library containing fragments carrying sample-specific barcodes. Rather than...is an author of 20+ scientific publications and patents.   ...

... important to know what kind of reagents you work with in experiments. Hence, I’m including this MSDS ...do a lot of genetic construction and cloning experiments. Not all of our lab enzymes are made by New England...AMINO can be beneficial for new researchers or students beginning to learn about biochemistry. The least...have a favorite lab app, please share it in the comments section below! Many thanks to our guest blogger...Pseudomonas aeruginosa. Follow him on twitter @100percentSDS.     Additional Resources on the Addgene Blog...

... none of the participants had preconceived opinions about “adeolitis.” The participants in my study were...used to stay up past my bedtime, poring over my grandparents’ medical textbooks by torchlight under the covers... affirm or challenge their beliefs. I asked participants to read short abstracts comparing the mortality...ideas about the controversial medical issues. Participants gave significantly more favourable ratings to...texts were otherwise identical. Likewise, when participants read an abstract for a study with findings that...

...collaboration - a sense of dialogue and community among participants, clarification - new and useful information,...them manageable. At Addgene, teams can add as many points as they like to the written agenda, but each only...and Accountability Agendas: Send around talking points for the meeting a day in advance. This gives everyone....  During the meeting, take notes on the agenda points. At Addgene, we like to do this live on the screen...for talking, not reading: Send around relevant documents, like proposals to review or project plans, before...send around a Google doc where people can make comments. At meetings, we just focus on the parts where...

...reported brightness measurements. Here, EGFP outperforms the improved folding variants, presumably due to.... A common requirement for live cell imaging experiments is the ability to follow multiple fluorescently... are mTagBFP2, EGFP or one of the improved GFP variants, mRuby2 or TagRFP-T, and an infrared fluorescent...In mammalian cells, one of the improved folding variants of EGFP like mEmerald or Clover is probably best...channels used for more abundant proteins or marking compartments. I hope this sheds some light on multicolor ...

.....we’re excited to help scientists share their reagents with the scientific community.   (click to enlarge...it ideal for CRISPR pooled library screening experiments. Find lentiCRISPR v2.(If you’ve peeked through... Zhang lab has shared over 40,000 CRISPR-Cas9 components over the years on their own or through Addgene...used as a negative control for mammalian RNAi experiments.  Find the scrambled shRNA vector. pLKO.1 - TRC...Tet-pLKO-puro - Tet-pLKO-puro contains all the necessary elements for packaging, reverse transcription, and integration...our depositors and requestors for making their reagents available and for supporting scientific sharing...

... to use the Gal4/UAS system, but the available reagents do not match your needs. If YFG is thrillingly...generate a transgenic fly There are a few important points to consider when choosing the vector you’d like...comparisons between lines expressing different variants of YFG because the transgenes are found in the...won’t be positioning effects). When designing experiments where you want to express multiple transgenes...chromosome your docking site is located). This amounts to a small difference of two or three weeks. 3....for. Making your transgenic fly and downstream experiments Once your gene of interest is cloned into a plasmid...

...response elements (TRE) in a promoter. Tetracycline and its derivatives serve as inducing agents to allow...inactive because a bound repressor protein actively prevents transcription. Once an inducer binds the repressor...turned ON. Types of inducible promoters Chemical agents, temperature, and light are all examples of factors...HSF-1), which subsequently binds to heat shock elements in the promoter, thereby activating transcription...various eukaryotic cell types (mammalian, yeast, plants, etc.) require unique promoters and there is very...

...diseases. They do that by studying patients to uncover new gene variants associated with conditions of interest...interest, then studying those variants in model systems: either human cells or mice. In a recent issue...implicated in disease or, in cases where particular variants or mutations have been reported in people, we .... You could use the kit to insert all of those variants into stem cells with the same genetic background...make induced pluripotent stem (iPS) cells from patients and try to compare those lines, they may all have...

...used together in the same experiment because the reagents used to detect the presence of an antibody depends...applications for antibodies You can use antibodies for experiments in a variety of fields including neuroscience...lysates, whole cells, or tissue samples. Many experiments that use antibodies use a primary antibody, which...buffer. Then, the purified protein can be used in experiments to compare the relative amount of protein present...such as Western blots or ELISAs, or for other experiments to study protein-protein interactions. Flow cytometry...2016) Aptamers, antibody scFv, and antibody Fab’ fragments: An overview and comparison of three of the most...most versatile biosensor biorecognition elements. Biosensors and Bioelectronics 85:32–45 . https://doi.org...

...experiment work? What are the various tools and components necessary to run it? And, most importantly, what...optogenetics: 1) electrophysiology and 2) behavioral experiments. I’m not an electrophysiologist so I cannot speak...any optogenetic experiment. There are numerous variants for the different types of opsins available and...these reviews for a discussion of the different variants [1, 2, 4]. And, of course, the best way to learn...learn about the many different opsin variants and how they can be used is to consult the many primary research...in which you monitor expression at various time points (e.g. every 1-2 weeks over 8 weeks) in order to...science considerations for your optogenetics experiments. Stay tuned!  Many thanks to our guest blogger...

...ChRger. In both experiments, neurons expressing ChRgers produced stronger photocurrents at low light intensity...invasive measures to perform in vivo optogenetic experiments. Intracranial injections of AAV are commonly ...properties best suited for less-invasive optogenetics experiments. The sequences in this library were designed ...in vivo minimally invasive optogenetics. Both experiments delivered ChRgers by intravenous injection of...the light was turned off.  Together these two experiments determined that ChRger2’s improved light sensitivity...

...ideal for human therapies and in vivo laboratory experiments ranging from transient gene expression in a particular...genome editing, optogenetics, and chemogenetics experiments. Recombinant AAVs (rAAVs) The primary application...available in rAAVs!   Figure 1: Diagram of AAV components      Cargo capacity Empty rAAV has a cargo...vectors with multiple genes packaged in them and use elements such as IRES or 2A to co-express them from one...take the next step – selecting your AAV and AAV components (serotype and promoter) for use in your experiment...Read Our Lentivirus FAQ Learn about Viral Vector Elements Resources on Addgene.org Read Our AAV Guide ...

...guide you through the basics of CombiGEM-CRISPR experiments. CombiGEM-CRISPR: The marriage of two simple ...interrogating their functions. The continual advancements in gRNA design necessary to achieve maximal ...highly flexible and can accommodate any genetic elements of interest. It can thus be tailored to address... the combinatorial expression of other genetic elements including transcription factors (Cheng et al.,...et al., 2015). Starting your CombiGEM-CRISPR experiments The first thing you will need is to get the list...should be verified by NGS before beginning your experiments. Figure 2: Strategy for assembling barcoded...

...from countless rounds of optimization. Dozens of variants exist with unique cutting features and evolved...engineering endeavors. For more information on Cas9 variants, check out our blog post on choosing a Cas9 enzyme...and is particularly convenient for targeting experiments requiring single strand DNA generation, such ... detection tool for rare, often hard to detect variants such as single nucleotide polymorphisms. Fast... the other Cas family members make for useful implements in the scientist toolbox. If your genome engineering.... Choosing the right Cas can really put your experiments Nsync.   References and resources   References...

...where to look for a coach. Many (most?) graduate students and postdocs may feel they have limited funds ...references and follow up with 3-4 to hear about those clients’ experiences. Good coaches will have loads of success...consider. Feel free to add other suggestions in the comments or email me with suggestions.  Most coaches have...through your PhD? Some coaches focus on graduate students. For example Dora Farkas, founder of Finish Your...with him.  He is also available for speaking engagements as are many of the coaches I have mentioned. ...Consulting works remotely, will travel for speaking engagements and has many areas of specialty from leadership...

...questions. Story telling shares many of the same elements as a STAR answer: a setting, a conflict, and a...which the story demonstrates I possess.   Bullet points on resumes are like cliff notes, so I use these.... This is a chance to highlight your prior accomplishments and demonstrate your science communication ...professional  First impressions matter, so make sure yours counts. Dress professionally but still like yourself. ...experience less stressful. First, you can make arrangements with those you live with to have a quiet place...talk. Some video conferencing software allows participants to “raise” their hand to ask a question or type...

... for a new generation of ABEs), consists of 3 components:  a deaminase (TadA, originally from E.coli,...modified ABE7.10 accordingly which resulted in variants named “ABE8.x”: “8” stands for the eighth round...x-d.  This resulted altogether in 40 new ABE8 variants. When comparing on-target DNA editing efficiency...PAM: NNGRRT) to create Sa-ABE8.x-m/d. With these variants, ABE8 could target many additional locations in...cells and return the corrected cells back to the patients. For example, defects in hemoglobin (𝛽-globin...continuous expression of fetal hemoglobin even in adult patients. In order to analyze the clinical use of ABE8 ...