... if the mutations are on the same chromosome. Homologous chromosomes recombine during meiosis, creating...homologous chromosomes in the parental generation, they are likely to stay on separate chromosomes in future...function when hemizygous, meaning that only one chromosome needs to contain the transgene in order to see...distance between the two loci of interest on the chromosome, the less likely crossing over is to occur between... which would get both mutations onto the same chromosome. Until crossing over occurs between the two loci...

... types of viruses generated in the lab. The four most common types of viruses grown in the lab are gamma...infection process (though it can integrate at chromosome 19q13.4 qtr. (AAVS1)). For these reasons, AAV...to work with BSL-2 materials, such as lentivirus. Most of this work will need to be done in a biosafety...a significant risk for aerosol contamination. As most centrifugation has to be done outside of a biosafety... and scrapes from sharp equipment are one of the most common forms of lab accidents, so extra precaution...significantly reduce the risk of an accident (the most common reason for lab accidents is not following...corners. For many researchers, viruses might be the most dangerous things they work with, and that can understandably...

..., Addgene had almost 18,000 CRISPR plasmids in our repository, and we have shared almost 300,000 CRISPR...plasmids. That’s almost a ninety-fold increase in size! From this collection, we’ve shared almost 300,000 plasmids... nucleases, the original CRISPR tool, remain the most popular deposited and distributed CRISPR plasmids...Addgene were knockout libraries, and they remain the most popular with our requestors. However, we can again...seem to never truly go out of style. Instead, an almost even mix of plasmids (by age) are requested and...

...FPs are fluorescent when ectopically expressed in most organisms. Furthermore, fusing FPs to another protein...used to study many biological questions. The two most common uses are: 1) to test the expression level...shift in wavelength from excitation to emission. For most FPs, Stokes shift is less than 50nm (often much ...environmental changes (e.g. pH, Ca2+ flux, etc). The most commonly used are GECIs - genetically encoded calcium...distinguish between closely related colors. Furthermore, most FPs  have a broad range of emission which will be... (EGFP) was optimized for 37°C, and is therefore most suited for mammalian or bacteria studies, whereas...or as long as 1 hour (mAmetrine1.2). However, for most FPs it is a few seconds to a few minutes. Photostability...

...choose to move away from academic science; there is most likely abundant opportunity to continue such discussions...discussing recent publications in a specific field(s). Most anyone who has spent any time in academia has likely...meeting takes place to ensure success. One of the most important is letting people know it’s happening.... be tailored to the group and format but should almost universally include the questions “Would you attend...discussed topic, and level of general busy-ness will most likely the primary drivers of frequency. At Addgene...start with one per month. When to call it a day The most ineffective type of meeting is the one that no one... should be well planned and carefully organized. Most importantly, they should be regularly assessed for...

...thinking about what comes next. To that end, one of the most important things that you can do now is diversify...landing my own position! Conferences One of the single most important things you can do to plan for your career...working professionals, with the added benefit that most people at the fair will probably be willing to accept... it stand out with key words and by putting your most relevant experience at the top. You may also consider...Here you can search for jobs, post your CV, and, most importantly, manage your professional network. This...hope to have, you must think outside of the box (most likely your lab, in this case). It is important ...Resources http://www.businessinsider.com/how-to-get-the-most-out-of-job-fairs-2014-3 https://www.forbes.com/sites...

...applications - now let’s dive right in! You know qPCR is most commonly used to measure relative quantification...correctly is imperative. Picking your chemistry  The two most common qPCR chemistries are hydrolysis probes (which...of pipettes and pipetting methods to find what’s most consistent in your hands.  When using multichannels.... They are often included in commercial sets and most machines have a protocol that automatically performs...performs the normalization step.  Machines Most general-use protocols, unless otherwise specified, were ...number of methods available for analyzing qPCR data, most with plug-and-play Excel spreadsheets available ... your analysis method. ΔΔCT/CQ method One of the most commonly used methods is the ΔΔCT/CQ method, which...

...However, when I enquired about roles in this field, most jobs needed at least 3 months experience working... practicalities of working in the field with the most basic equipment and was a tremendously positive ... that all commercial companies are like this, as most have a more stable funding situation and level of... of work than research or academia, and so most positions in industry are more stable than this. However...experience is that the technology is so advanced that most of the procedures in the lab are done with robots... the phone or explain how various plasmids work. Most people who call or email me are scientists who are...self-motivated, because you need to work independently most of the time and plan your own work schedule.  I ...

...sites on different chromosomes in living cells, the system requires high-copy chromosome-specific loci in...study the spatial and temporal arrangement of chromosomes and how these arrangements may affect nuclear...based multicolor labeling system to visualize chromosomal loci in human cells (Ma et al., 2015). To do ...inter-locus distances. They tested this system out on chromosome 19 by targeting 46 repetitive loci that contained...Hanhui et al. “Multicolor CRISPR labeling of chromosomal loci in human cells.” Proceedings of the National...

...the C terminal of the tag. Of the most common tags, FLAG is the most charged, with 5 negatively charged...and applications. In this blog, we will review the most common tags, the antibodies used to detect them,...you access to a variety of antibodies that have almost certainly already been optimized for your application...specific monoclonal antibodies. Below we review the most commonly available tags and their antibodies along...antibody to Myc was developed in 1985 and is still the most prevalent clone used today! Goat, chicken, mouse...

... this here and now. Our closure was earlier than most companies. Since the start of this crisis, we’ve...mobilized on COVID-19 research, cures, and diagnostics. Almost immediately we began to receive urgent requests...guidance. We were also dealing with pending orders, but most labs around the world were closed and not receiving...challenge was our Finance Team. With Addgene closed for almost two weeks and with labs worldwide closing for unknown...Protection Program loan and received enough funds to make most of our payroll for the spring and early summer months...distributes and fulfills requests for scientists doing almost every type of biomedical science research. Our ...

... next steps in the process involve repairing chromosomal DSBs. It is important to know that cells possess...extensive DNA synthesis. NHEJ also finishes repair of most types of breaks in tens of minutes – an order of...should possess indels in the majority of their chromosomes within a day. Another factor expected to impact...or more can simultaneously introduce a pair of chromosome breaks, and could result in deletion of the intervening...Stark JM (2009) Limiting the Persistence of a Chromosome Break Diminishes Its Mutagenic Potential. PLoS...EA, Giovannangeli C, Jasin M, Brunet E (2014) Chromosomal Translocations in Human Cells Are Generated by...

... proteins, etc., that you are interested in. The most obvious component of a biosensor is the thing that... culture and animal models primarily. Fun fact: Almost all of the biosensors discussed are genetically... Within these ions, calcium is by far one of the most popular, with dozens of sensors developed for it...potentials; it’s an ion of interest for many disciplines! Most of the sensors designed for calcium generate a fluorescent...intracellular, extracellular, and even the ADP:ATP ratio. Most of these sensors are fluorescence-based and have...tips. Still worried? We make virus with some of our most popular biosensors already packaged and ready to...

...and is found mostly in blood. IgD While IgD is coexpressed with IgM on the surface of most immature B cells...blood, where it’s the second most common antibody after IgG; but it's the most prevalent antibody in secretions...valency of two and has no subclasses. IgG IgG is the most abundant isotype in blood, but it’s also found in...where it protects mucosal membranes. This isotype is most commonly a dimer with a valency of four. There are...

...Harvard Medical School working on human artificial chromosomes. Montreal, QC, Canada 45.5017°N 73.5673°W – Vanessa...to full reproductive health care. And it’s needed most in low-income and minority communities, where these... hardest. According to the Guttmacher Institute, most women of reproductive age live in a state with at...about my work because it's important to our health.  Most people won't argue with that idea, but they might...into a successful PhD thesis. We use the same atmospheric physics to understand exoplanets that we use ...organized the march was, as well as the friendly atmosphere that was created by the many thousands of science...science, but I worried (and continue to worry) that most who participated will consider that marching in ...

...Microscopy Fluorescent Widefield Microscopy is the most common and simplest form of fluorescent microscopy...developed that allow for sub-diffraction limit imaging. Most often, these techniques provide a 2-10x improvement...interference from blurry out-of-focus light. The most difficult characteristic to capture in these types...resolution on the millisecond scale may be needed. Most commonly these experiments are performed on a fluorescent... a fast, sensitive detector such as a scientific CMOS camera. If plasma membrane dynamics are the main...small model embryos >15 μm Thsee are some of the most challenging samples to image as they require both...quality images for this type of sample as they are most efficient at excluding out of focus light. However...

...stream to commute to and invade multiple organs, and most importantly escape the immune mechanisms of the ...known as the tumor microenvironment - a vital atmosphere created by the cancer cells to shield themselves...cell-surface tolerance-producing proteins are one of the most commonly used immunotherapeutic agents. Currently...receptors (CARs) on their surfaces have been the most acclaimed form of immunotherapy in recent times.... validation to identify populations that are the most likely to be positively impacted by the trial. During...potential beneficiaries. Collaborative therapy For the most efficient outcomes, collaborative therapies must...questions in this exciting field of research. One of the most important: How will cancers evolve and mutate based...

...email me with suggestions.  Most coaches have websites you can review and most create useful online content...and do not know where to look for a coach. Many (most?) graduate students and postdocs may feel they have... have limited funds to invest in this. However most coaches who focus on scientists are sensitive to this...effective and also help keep the coaching costs low. Most will work with you remotely (by phone, Skype or .... This larger shop might appeal to some although most scientist coaches can help with transitioning out...

...delivery and gene therapy and represent one of the most studied viruses. Adenoviruses are non-enveloped ...isogenic AdVs encoding TALENs targeting the same chromosomal region. The CRISPR/Cas9-derived RNA-guided nuclease-induced... will interact with sporadic double-stranded chromosomal DNA breaks “that always happen naturally.” These...Muscular Dystrophy, Alpha1-antitrypsin deficiency (most widely used so far) or for cancer gene therapy such...Current state and clinical perspectives. Biochemistry Moscow 81:700–708 . https://doi.org/10.1134/s0006297916070063...Ventura A (2014) In vivo engineering of oncogenic chromosomal rearrangements with the CRISPR/Cas9 system. Nature...

...efficiency than in iPSCs, but with the caveat of mosaicism. To circumvent this problem, they co-injected ...Indeed, they observed that 41/42 embryos were non-mosaic at 3 days post fertilization. Since Ma et al. designed...term ‘editing.’ While repair using endogenous chromosomal DNA is appealing for heterozygous mutations, ...needed to define whether off-target editing or mosaicism would occur. Although Ma et al. looked at various...this detailed analysis is not possible with IVF, as most of the cells are needed for proper development. .... Ma et al.’s experiments showed a 3% mosaic rate, and this rate would need to be much, much lower before...