...distributed in digital or analog media formats. Cognitively demanding format. Equipment, expense and expertise...with is "how?" How are you going convey your information to your audience? There's so many ways options...incredibly varied in how they prefer to consume information. Personally, I would much rather read the instructions... science.                    Two of the platforms we use at Addgene: YouTube, for longer educational...octogenarians is going to be a hard sell!) Digital platforms often have demographic data of their users readily...geographic location. For example, not all social media platforms are available or used in all countries; not all...mitosis.   Consider also the amount and type of information you want to convey. Small tidbits and fast/fun...

...provides summary information on Class 2 Cas enzymes. The information is organized in wiki format and includes...display information for a given Cas (SpyCas9a) from CasPEDIA.   Helpful resources and information For ...could arise where you still need more information. If you are performing a viral delivery of Cas, the size...the features? CasPEDIA has collected relevant information on all the Cas enzymes you know and love, and... heard of. Each entry displays the following information for a Cas enzyme: Nuclease properties (CasID...properties of Cas enzymes. That’s a lot of helpful information! To get you started, we’ll give a quick overview... substrate (nick, bind, cleave, mixed). This information represents the ‘big picture’ features of the ...

...More concretely, scientists gather information about reality informing people and their governments on what...article are those of the author do not represent a formal stance taken by Addgene or its staff. In Washington...reality and science is how we learn about reality. Formally, a government can be defined as a social contract...into in order to protect their rights (2). Less formally, governments are the organizations that make and...other citizens. Thus civilization proceeds in an informed fashion. For science to be useful to governments... of the bargain and do the research required to inform the public (8–13). Facing the possibility that ...march include: General inclusion of science to inform policy (17). The facts surrounding a matter or ...

...Arabidopsis seed following floral dip transformation next to an untransformed seed from the same plant beginning...Selectable transgene markers for plant transformation fall into two main categories: antibiotic/herbicide...resistance genes usually involves killing the untransformed seedlings or callus (undifferentiated plant ...For example, with CRISPR-Cas9 gene editing, transformation may be identified by resistance to an antibiotic...antibiotic. Successful editing of transformants then typically occurs in T0 plants (from callus) or T1 plants...contaminate it. I have used it for selecting transformation events and am currently planning to use it ...beginning to germinate. (Top right, second panel) Transformed seedlings can be selected without antibiotics...

...procedure to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As...false positive colonies. Transformation - the linear fragments are transformed into E. coli and plasmid...important is the transformation efficiency of the strain: when asking the cell to perform multiple recombination...assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation. However, while...adopted by any laboratory. All procedures are performed using the same three step protocol:  Fragment... encode homology arms to allow immediate co-transformation with linearized vector backbone. DpnI digestion...competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications will require ultracompetent...

...bacterial transformation One protocol that has been automated is high throughput transformation. With our...a couple of platforms that fit 48 well q- trays allowing us to do up to 96 transformations in one run ...easier and more efficient. Here, I’ll discuss transformation, DNA purification, and filling vials with agar...deposits (over 100 deposits per month!), manual transformations were taking a lot of time. Enter the Biomek... We wanted a machine that would improve our transformation protocol and, after looking at a few options...run compared to doing about 50 transformations in the same amount of time by hand! Setting up a basic ... fine piece of machinery we have been able to transform 96 samples in about 4 hours with a greater than...

...anyone who has ever worked to sequence DNA on NGS platforms will tell you, while the process has become much...sequencing. This process is completed in a plate format yielding anywhere from two to six plates of 96 ...donated MiSeq.  For Complete Plasmid Sequencing we perform a 2x251 run on our MiSeq that takes about two days...involves a significant amount of QC. First, we perform Picogreen quantification on each and every isolated...samples for Viral Genome Sequencing (VGS). QC is performed throughout the library preparation process and...Aligning the sequence The first step we take when performing quality control of newly deposited plasmids is...detecting common features using Snapgene. If we find information that differs from what we would expect given ...

...purification before transformation. After transforming cells, you should perform colony PCR with vector..., ligation, and transformation. While restriction cloning is still routinely performed by many labs, a...products are then transformed directly into chemically competent E. coli and transformants selected with ...+ + Gel Purification + - Ligation + - Transformation + + The FastCloning process in brief In ...for this procedure to work properly. Directly transform chemically competent cells with the digested mixture...-cycle PCR leads to the most colonies after transformation. When a 1:1 vector:insert ratio is used in ... and 2 µL of digest product are used in the transformation, >70% of the colonies are correct. Good luck...

...AMA   Guidelines for giving feedback Use formal and informal mechanisms for feedback (more on this below...her management style. Formalizing the feedback session Giving regular, informal feedback is a very effective...Using this standard format not only enables you to discuss a person's performance or what has been learned...scientists seem to find it easier to give feedback in formal sessions using a specific list of questions after... it helps to plan effective next steps. For a formalized feedback session, answer the questions below ...discuss openly. Both parties get used to using this format and it becomes even more effective over time if...of indirect language You must be honest about performance problems. While giving corrective feedback isn...

...organizational performance and often have to be able to quickly synthesize information from a variety ... their visual format is quite appealing to many audiences. While all of these platforms were developing...not hosted on the platform, they can be hosted on YouTube, Vimeo, and other platforms (such as university...quickly. Taking what I loved from each of these platforms (such as social networking, visual appeal, instant... across multiple other sites), I created a new platform for sharing video abstracts about research: www.WeShareScience.org...www.WeShareScience.org. WeShareScience is a free platform for sharing short, 3 to 5 minute videos about ...university websites). WeShareScience is a social platform that provides links to the videos, keeping the copyright...

...together through the transformation/replication process. LIC employs long overhangs to form a stable association...now ready for transformation. Use 1-2 μl of annealing reaction for each transformation, following a standard...association between fragments, allowing for transformation without ligation. Because of its dual polymerase...standard polymerase protocol), causing the enzyme to perform exonuclease activity until the first "G" in the...sterile dH20 to 40 μl Step 4: Amplify Insert by PCR Perform PCR amplification of your insert following the ...heating to 75° for 20 minutes. Step 6: Anneal and Transform Mix your treated vector and insert at a molar ...

...The ability of our users to discover all this information, however, did not improve at the same pace as...as the quantity and quality of the information. As a result, resources on the website were hard to find... than that: the general “findability” of the information on the site. Different users have different strategies...paths, the organization and completeness of our information, and the accuracy and relevance of our search...the advantage of a very specialized domain of information so we were able to fine tune our approach much...than if we were dealing with general purpose information. Below are a few of the strategies we used in...Categories We created a categorization of our information in order to more granularly customize the search...

...optimization as each antibody performs differently and some antibodies impact the performance of other antibodies...beautiful results. Performing an immunofluorescence experiment The process to perform IF can be broadly...important considerations to make when planning and performing an IF experiment. What is immunofluorescence... different species and colors makes it easy to perform multiplexed IF. By using multiple primary antibodies...in IF experiments. Precisely which controls you perform will depend on your experimental context.  Choosing... to use, then the primary antibody will likely perform as expected in both.  How do you find validated...may be, it is always important to verify their performance in your hands and in your specific experiment...

...80) are transformed to 100. This transformation (illustrated in the graph below) is performed because ...order reagents Perform your wet lab experiment Analyze the data For more detailed information on using CrispyCrunch...even more.) However, none of the tools we found performed batch guide design, batch primer design or batch... burden of having to manage all the standard information that goes into a 96-well plate and comes out ... reports. With CrispyCrunch, you get the all information from Crispresso plus a summary per batch. Ranking... at a genome by eye. We came up with a ranking formula that does just this: Rank = (compressed CFD score...the DNA target. CFD scores are then modified (“transformed”) such that weak interactions (e.g., scores <...

... base editing platforms for plant systems.  Figure 2: Plant base editor platforms and their molecular...72 hours post-transformation/transfection in a transient assay. For stable transformation, regenerated ... reducing the likelihood of detrimental indel formation. After binding with DNA, the CBE deaminates a ...G)? You may wonder if a CRISPR-base editor can perform this change. However, there are no naturally known...deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring the defective CamR ...genome edited plants via Agrobacterium-mediated transformation. We also constructed smaller vectors suitable...suitable for transient assays using protoplast transformation. Cytosine base editors for plant There are several...

...to read a formulaic fiction story that doesn’t require any detailed retention of information or formulation... extraneous cognitive load refers to the way information or tasks are presented to a learner; and germane... particular way of organizing and connecting information and a logic to the types and timing of questions...question constantly (and so should you!)  Amount of information and level of detail Again, this is an obvious...load, than certainly amount of attention and information input. If you’re reading a book, you only have...quickly they can understand and process your information. A class of three year olds is unlikely to be...I would think about how I was presenting the information. Maybe I needed to remove a distraction, such...

...copies of your predicted sequence information. Collect plasmid information in a binder or digital notebook...barcoded tubes and plates. The critical pieces of information for a given plasmid are the backbone, inserted..., the predicted full plasmid sequence. Other information may also be critical to the maintenance and use... with us, it will be easy to find all of the information needed for the process. Always sequence verify... Once the physical sample and its associated information are at Addgene, we are ready to sequence verify...always be derived from a single colony to ensure uniformity of the culture. Sequencing directly from the ...concentration, and not all providers are equipped to perform Sanger sequencing from glycerol stocks, so make...

...feedback in multiple formats      Ask for input and opinions in meetings, via email, at formal communications... hand and eagerness to get, as well as give, information to make improvements. Members connect directly... this means encouraging a culture of maximum information flow so everyone knows what is going on and can...break, go exploring outside the team, and bring information back. No team has all the answers – reinventing...frequently One of the most important paths of information flow is the one that brings feedback back to ...from at least some employees. Watch out for "information allies" – these are employees in your group that...communications events or at informal gatherings, like at lunch. Many experts recommend using anonymous surveys with...

...pooled libraries have allowed scientists to easily perform genome-wide screens to effectively and efficiently...sgRNAs per gene. These libraries provide more information while using less resources and are particularly...that improved sgRNA activity and thus library performance. The Root and Doench lab’s optimized genome-wide...CRISPRi, and CRISPRa libraries have been shown to outperform other popular CRISPR libraries (see below). Let...colon cancer) cells and assessed the libraries performance via the depletion of a gold standard set of genes...developed sgRNA-level metrics to assess library performance in viability screens. Based on these metrics ...sgRNAs, Brunello even with only a single sgRNA outperformed another CRISPR library with 6 sgRNAS, highlighting...

...found that many researchers feel that they lack formal training in a variety of transferable skills. At...many graduate and post-doctoral programs lack any formal or mandatory professional development training....you should have a business card; it’s a bit more informal, and a lot easier to hand out than a CV or resume...the connection you made at the job fair when you formally apply for the position. This post on Business ...call, make sure to get that person’s contact information and again, make a personal connection! Even if...are, and who they know all in one place! This information can be critical to a focused and effective job...probably have a lot of relevant experience and information to offer. However, if you hope to have a career...