We narrowed to 928 results for: Mpl

... Their trust in the quality of the content is implicitly defined by their trust in the piñata builders...deals in post-experiment quality control, a good example is CRISPR-GA. These tools assess repair events ...published 1 Jan 2013, no longer available), is a simple catalogue of reported sgRNA designs. The second...similar to EENdb than CrisprGE, but is also a good example of how database tools can be used to aid design...this category are the most common between tools. Examples: “single-target design”, “multi-target design”...considered extremely useful, depending on design goals. Examples:  “feature aware”, “SNP aware”, “secondary structure...post-design process such as primer and plasmid design. Examples:  “multiplex design”, “multi-method design”, and...

... Many of these examples we’ve done annually - they are fan favorites! 6. Keep it simple This may be an...fun into the mix. Thus, our founders and early employees spent this remaining time by elaborately pranking...University Park at MIT.  I’m sure at some companies, employees wouldn’t be caught dead coming into the work vicinity...their teammates and project collaborators have completed their work thoroughly. It’s kind of a chicken ...seems to be a balance of productive and happy employees. The teambuilding culture helps us build relationships...continue the tradition and plan enjoyable events for employees. We now have a team of three (myself included)... best events have been at the suggestion of an employee, or have come to fruition from a lunchroom conversation...

... (5,12,13). For example, in the Ligase Detection Reaction (LDR), probes complementary to the sequence ...with a different complement strand covering all 16 possible NN pairs in the template. The probes were ...ligation of fully base-paired nicks with no gaps. For example, DNA assembly methods, such as Gibson Assembly®...without the exo/polymerase generation of annealed complementary regions. Why use a high fidelity ligase? Numerous... annealed to target DNA (genomic DNA or a PCR amplified fragment) and treated with a high fidelity thermostable...rounds of melting and annealing allows linear amplification of the probe ligation product. With the right...uses two sets of probes to achieve exponential amplification. What does fidelity mean for a ligase? DNA ligases...

...the best choice for small sample sizes, small regions of interest, or complex sequences such as G-C rich...However, Sanger sequencing remains useful for small sample sizes and shorter regions due in part to its low...high accuracy. Conversely, you can only run one sample at a time, which can become costly and impractical...the bases (and, if necessary, separate different samples), the resulting data, or reads, are essentially...lot of puzzle pieces. An assembly algorithm is employed to put these puzzle pieces together. Once they...control. NGS is generally cheaper if you have many samples and can output more data, and it typically results... regions or for higher accuracy of a particular sample.  Long-read sequencing (“Third-generation sequencing...

...you are shuttling between. For example, if you are using E. coli simply for easy cloning and quick DNA...function in unrelated host species, even from completely different kingdoms of life. That’s convenient... for researchers who want to do experiments in complex eukaryotic cells, but would prefer to grow, clone...into plant genomes. Eukaryotes are a bit more complicated. Yeast vectors may contain an autonomously replicating...episomal vectors is to include viral oris to drive amplification, which helps increase cellular uptake, but this...systems don’t always translate to eukaryotes. For example, beta-lactam antibiotics like penicillin and ampicillin...pathways shared by bacteria and eukaryotes. For example, puromycin inhibits protein synthesis across kingdoms...

...people outside the department to accomplish a variety of goals. For example, I worked with some microscopy...experts that complement your (scientific) skill set and is therefore very useful for complex projects. It...collaborations are becoming increasingly important as more complex approaches to scientific questions become popular...mutual benefit.  Our partnership with SnapGene, for example, allows us to display high-quality maps and annotations...partner, connect the relevant teams, coordinate implementation, and now must focus on managing the established... in teamwork and collaboration management. For example, you may already be part of a scientific collaboration...common interest so naturally this may be easier to accomplish with people at your organization. You can meet...

... encode which amino acids (see example to the right). With such simple rules, you might think it’s easy...express proteins in heterologous hosts. If you simply amplify a gene of interest from the human genome, for...varied codon preferences among organisms aren’t completely understood, but some possible reasons include...resources for the entire translation process. For example, during high growth rate conditions, E. coli preferentially...encoded by highly abundant, charged tRNAs. For example Tuller et al. found that translation efficiency...translated regions a chance to fold properly. For example Pechmann and Frydman found that tracts of non-optimal...producing and charging specific tRNA pools. For example, tRNAs used in genes encoding amino acid biosynthetic...

...fully summarized in a short blog post, but some examples I’m personally interested in include: intracranial... preference.   Historically, ICSS was done by implanting an electrode and non-specifically electrically... also models reinforcement but is considerably simpler than ICSS. The test chamber is divided into two...expression of the early response gene c-fos. For example, using Chr2 stimulation should result in robust...Inhibition of neuronal activation is a little more complicated since basal neuronal activity in your brain region...c-fos expression to get the evidence you need. For example, as shown in this study [3], hippocampal neurons...but before you do any type of behavior analysis, simply take a few animals and stimulate them for 30min...

...you only need 1-2 ul of sample and the run time is just a few minutes per sample. During the run, DNA fragments...absorbance. Then you measure the absorbance of the DNA sample.  These absorbance measures give you an idea of...dilution factor x 50 ug/ml This method is quick and simple and doesn’t require any special reagents. However...especially when you expect low concentrations in your samples and are often used to quantify DNA for next generation...fluorescence-based methods require a standard curve, a set of samples with a known DNA quantity and their corresponding...That way, you can compare the fluorescence of your sample against this curve to quantify your DNA prep. Though...yourself as many fluorometers will calculate your sample concentration for you. Agarose gel electrophoresis...

...are a few examples. Make sure you use a proper plasmid (i.e. contains the α-complementation cloning MCS...isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds and inactivates the lac ...to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional enzyme...enzyme. This process is called α-complementation. The system described above was put to practical use in the... inserted it into a plasmid, creating an α-complementation cloning vector. When a cloning reaction goes...interrupted as shown in cell A, and thus will not complement the cell's β-galactosidase mutation. An unsuccessful...functional β-galactosidase enzyme through α-complementation (cell B). As shown on the representative plate...

... research is about and what you think you can accomplish. No matter how many times I say this, candidates...specific questions and experiments. Nobody EVER complains that a seminar was too easy to understand. Make... to a Perfect Science Talk." 4. The chalk talk, simple in principle The first question I usually get about... important to understand. (1-2 sentences). For example: Distinguishing correctly folded and misfolded ...essential for human health. Failure to do so is implicated in several human diseases. Proper patterning ...focus of your first grant? (1-2 sentences) Simply stated, what are your aims? You could directly ...official aims, but I suggest distilling them into simple goals/outcomes. This isn't the time to go into ...

...are more techniques and strategies one can employ beyond simply incubating cells with single-fluorophore-conjugated...ELISAs.  A popular method for indirect labeling employs a primary epitope-targeting antibody that is conjugated...antibody can originate from a range of species; for example, rabbit or mouse. In a second step, a fluorophore-conjugated...interested in one of the various cell populations; for example, CD8+ T (“T killer”) cells. Typically, you would...FACS achieves greater purity of your sorted cell sample than MACS. An effective strategy to sort out your... B cells, and dendritic cells. In this case, we simply need to label the lineage markers of all cell types...selected color will not be CD8+ T cells and we can simply “dump” them by gating on the negative population...

...fluorescently labeled probe. If the DNA is complementary to the probe (for example, has the sequence of the mutated...searching for), then the probe will bind to the sample thereby labeling it. The label can then be visualized...strength of the signal in comparison to a control sample. Southern blotting applications range from confirming...experiments to forensics and clinical diagnostics. One example of southern blotting being used to identify a gene... MstII and southern blotting are conducted on a sample from an individual without sickle cell anemia, ...interest by recording DNA abundance throughout the amplification process, rather than just at the end as in standard...DNA sequences to see which is more abundant in a sample. Whereas Southern blotting is labor intensive and...

...an account of the material resources needed to accomplish your science, and can include: equipment, reagents...then start identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline...consumables (such as pipet tips, glassware, etc), get a simple list from your current P.I., lab manager, or administrator...need to tap for your future research team. For example, if you’re going to need a tech because you’ll ...regions where the host institute is located. For example, salaries in Kansas versus salaries in NYC will... most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE... that have the means to support your work. For example, if your work requires specialized equipment, try...

...tissues, cells, plants, fungi, or bacteria, keep samples cold and work quickly to mitigate RNA degradation...180°C for at least 4 hours.   If your final RNA sample(s) are resuspended in water or TE buffer, store...freezer.   RNA extraction methods evolved into a simple protocol still used today There are many alternative...for RNA extraction and purification. There is one simple method that works, and variations to that method...requires a lot of time and depending on how many samples you may have, access to one or more large, expensive...effectively separated from DNA and proteins by a simple extraction protocol with guanidinium thiocyanate-phenol-chloroform...thiocyanate-phenol-chloroform. In this method, samples are still homogenized and lysed in a guanidinium thiocyanate...

...important that the digest go until completion. The time required for complete digestion varies for different...between samples. In addition to a DNA ladder standard, it is also a good idea to run an uncut sample of each...plasmid to the new fragment thereby generating a complete, circular plasmid that can be easily maintained... gene insert with respect to the promoter. For example, let’s say your plasmid backbone looks like the...    Alternatively, this whole process can be completed using a single enzyme if your insert is flanked...you to either stain your gel after you run your samples or add the stain as the gel is being made (post... digestion to anneal to one another and form a complete, circular plasmid. You then add DNA ligase to ...

... used immediately for amplification.*Pro-Tip*: The cells will become completely confluent and the media...particles = number of particles per mL of sample. The purity of the sample can be estimated by looking at the...negligible toxicity. However, their production is more complex and the presence of helper virus contaminants have...AdV can be found in reference 3. Production, amplification, and quality control of 2nd generation AdV   ...bacterial strain not prone to recombination for amplification. The final purified plasmid stock should be ...incubation as it will result in very low titer. Amplification (1-2 weeks) - The rAdV-S is used to infect more...and clump together ~3 days post infection. The amplified virus can be harvested once roughly 50% of the...

...your sample to. For example, we use our negative control to compare exposure times of our samples. A prominent...cannot be reproduced and projects that cannot be completed. A 2015 review analyzing and explaining the antibody...Baker, 2015), causing a huge waste of precious samples, time, and funds – up to US$350 million in 2015...techniques on rodent brains and is expanding to more complex brain systems such as non-human primates and humans...noise. We then compare the negative control to a sample with the primary included to confirm that the signal...antibody and not from the background noise. If the sample images similarly to the negative control, that ...weak or with general non-specific binding. If the sample differs from the negative control, such as showing...

... CRISPR samples. We invite you to jump in and try it out, or take a look at our live examples: experiment... principles listed here. It employs biological best practices. For example, it ranks guides by CFD score...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At...score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you control over your...if desired. It works as a whole or in part. For example, you can analyze experiments designed outside CrispyCrunch...with NGS data were: Crispresso CrispRVariants AmpliCan We chose to adapt Crispresso by Luca Pinello ...active guide. With these constraints in mind, we implemented the following algorithm for optimal mutation ...

... as the building block of our understanding of complex biological processes. Many tools exist to perturb...in the context of the biological question. For example, while gene knockouts have long been considered...silencing,” of any gene that contains sequence complementary to the dsRNA (Wilson and Doudna, 2013). RNAi... translation, or chromatin modifications of complementary gene sequences (Figure 1). As a result, gene...unmodified dsRNA elicits immunogenic reactions that complicate the effectiveness and interpretation of RNAi.... of interfering RNAs.   RNAi vs. CRISPR: Complementary techniques It is fair to say that CRISPR has...of the technique—all that is required dsRNA complementary to your gene or genes of interest to trigger...