...using primers that contain the restriction sites needed. Functional restriction sites must be a certain...definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus. Third...

...will need compensation? To know if you will need to compensate in your flow panel, you will need to know...would say that APC is “bleeding,” or spilling over, into APC-Cy7 channel and needs to be compensated out...this bleed. A compensation value of 3.7 for APC into APC-Cy7 applied next, which fixed the bleed in this...fluorescent markers are read by the same laser, they will need to be compensated for one another. This overlap ...fluorophores FITC and PE on the same laser, you will need to compensate between the two of them because they...reads them out on different lasers, you should not need to compensate between them. Another rule of thumb...of the fluorescent markers are similar, you will need to compensate between them (although there are exceptions...

...Identify the skills needed to use these tools Outline the knowledge and skill sets needed to do your science...that you’ll need to tap for your future research team. For example, if you’re going to need a tech because...Experiment Designs Consulting, Inc. “How much do you need [for your lab start up]?” A hiring chair or dean...doesn’t have to be this way, if you know what you need. Most often, early career scientists aren’t trained...instead have our clients focus on their experimental needs and consider a budget as a resource checklist. A... budget is an account of the material resources needed to accomplish your science, and can include: equipment...discussion first defines the material resources needed, then determines how much those resources will ...

... germline Selecting for seeds that have the tDNA insertions (usually via seed growth on antibiotic-containing...fungal growth. Healthy plants produce lots of seeds. More seeds mean more chances to recover a transformant...of time to produce seeds after you expose the flowers to Agrobacterium. More seeds mean more chances to...produce lots of seeds. I typically use 8-10 plants per construct I transform. More seeds mean more chances... transformants based on expression of RFP in the seed (Emami, Yee and Dinneny, 2013). Preparing plants...transformation efficiency. If you see many mature siliques (seed pods), your plants may be too old to transform. ...mature prior to transformation are just full of seeds that you know won’t harbor your transgene. You can...

...timely feedback. We often associate feedback with end-of-the-year performance review, but feedback should...assume that reports want to succeed at work and providing feedback is needed for that. In many cases reports... for the feedback, but at least you can start making changes for the best. Positive feedback is equally...you team feel safe. Your team members need to trust you and you need to encourage them to speak their minds... and by offering advice on areas where you might need some help. Today in our transferable skills guide...! I was a poor delegator at first. I knew what needed to be done and how to prioritize but I  just added... is no way around the fact that new team members need to be trained, which may seem like a waste of your...

...that can’t set seeds. I study the development of the mutant plants, but the seeds needed for experiments...the green seedlings that don’t have the transgene and save the red seedlings to collect seed for further...RUBY-expressing Arabidopsis seed following floral dip transformation next to an untransformed seed from the same plant...genes usually involves killing the untransformed seedlings or callus (undifferentiated plant cell culture...dip). In the next generation the CRISPR construct needs to be segregated away to prevent continued production...simplify the identification of these mutants will speed up my work and make it easier to study these mutants... plants will be red and will be able to produce seeds. However, they will not be useful to study because...

...you can optimize or troubleshoot a western, you’ll need to be familiar with running a gradient, which will...proteins are nuclear- or DNA-binding proteins, you may need to sonicate your lysates in order to release the...optimal gel for your experiment.   Ladders If you need very accurate protein measurements, consider using...visualize them with a conjugated antibody. If you do not need such precise measurements, you can use a colorimetric...left-hand corner with a pencil (not a pen — they bleed!), cut off the upper left-hand corner, or place ...troubleshooting step to take. If you’re not sure if you need a smaller pore on your membrane, run a double-membrane...stick to the membrane by stripping away the SDS. To speed up larger proteins and prevent them from under-transferring...

...materials, you’ll need to use aerosol-tight caps, compatible with your centrifugation speed, to cover the ...plasmid (packaging plasmid). Viral production then proceeds by transfection of all three plasmids into a producer...proto-oncogenes, which can lead to cancer. These viruses need to be handled in a BSL-2 (or BSL-2+) lab and require... your lab coats and double gloving, so that, if needed, a layer can quickly be removed and disposed of...materials, such as lentivirus. Most of this work will need to be done in a biosafety cabinet. Many researchers...to be done outside of a biosafety cabinet, you’ll need to take extra steps to ensure that particles are.... Finally, when spinning down lentivirus, you’ll need to use centrifuge bin covers, which you should open...

...of interest. You can also just follow feeds curated by Meta. Feeds automatically update as papers are published...personalized feed is simple. You can enter as many search terms as you like, and your feed will update ...your feed which means only papers than contain all of your search terms will show up in your feed. Feeds...score, is also displayed in your feed. By clicking on a paper in your feed you can see suggested related ...characters! Using Twitter to find papers  You’ll need to start following relevant accounts in order to...search engine. Instead, you create your own research feed by entering keywords, concepts, journals, and/or... to help you expand your search. Results in your feed are ranked based on their influence on the network...

...method to another or change primary antibodies as needed. However, more antibodies mean more optimization...can be more difficult to source, especially if you need a specific conjugate, and changing your reporter...preparation Before you can blot your proteins, you’ll need to lyse and denature them. If you are intentionally...you are quantifying your western blot data, you’ll need to determine the linear detection range of your ...to the other.  Type of gel The type of gel you’ll need depends mostly on the weight of the proteins you... each other. Once you know your gel type, you’ll need to pick the percentage. You can get a single-percentage... 4–20% gradient   Handcast or premade If you need a gradient gel, the decision is probably already...

...exactly your lab needs from an ELN (the full article is available here):   Does your lab need to keep paper...exported or printed from the ELN when needed. Whether it is because you need to keep paper records as well as...to do it when needed. How many people from your lab would use the ELN i.e. do you need a system that ...research? Electronic lab notebooks and researcher's needs The familiar paper-based lab notebook is still a...institutions tend to think about the long term, so they need a solution for which they are sure that there will...electronic lab notebook starts with defining your lab’s needs The values of ELNs show in the long run as they ...currently exchanging information with your team? If you need to exchange information regarding lab work with ...

... if needed. And since we live in the digital age, make sure to research any software you may need: cost...possible COVID-driven virtual drain. Audience needs high speed internet access. High probability of real-... of any given country have equal access to high-speed internet. Before you begin any work, make sure you...produce an outcome you’re happy with. Know if you’ll need specialized equipment and/or expertise from the ...to tell a compelling story in an image or do you need a full exhibit? In a five minute activity at a street...demanding format. Equipment, expense and expertise needed can increase exponentially with complexity. ... and physically. Easy to incorporate visuals if needed. Traditional media formats are quite competitive...

...just as easily discover you need to add some people if you find there’s a need for an expertise your current... and by offering advice on areas where you might need some help. Today: Teamwork. My first experience ...may be relatively permanent; some teams may only need to exist for a year, or a few months, or even a ...project that never ends is a poorly planned project indeed. Mission team A Mission Team is assembled to make...requires a broad knowledge base is going to necessarily need a diverse group of people to provide that base. ... out the door much faster, but getting helpful feedback from other people who have a stake in the outcome... Social interaction is a good thing Part of you needs to interact with other people, whether you want ...

...additional data needed to rule out alternative models or to strengthen your new model?  PFs need to be regularly...your postdoctoral experience, define what will be needed to achieve your intended next career stage and ...competitive for a Group Leader position, what will you need? Multiple first or corresponding author publications... you get there and more practically, when do you need to be there? A typical biomedical postdoc fellowship...Group Leader position, key first author manuscripts need to be ready to write by Year 3.5. Thus, it is critical... directions of the lab, gives you constructive feedback on your research, and teaches you how to write...mentor is. If the person makes time for you, as needed, this is a strong indicator that this person cares...

...that we chose needed to fulfill certain requirements: Customizable to our specific needs Modern and well-supported...usage of the scripts needs to be customized in the pipeline for the specific needs of the user. Thus, a... repository continues to expand, we realized the need to modernize our approach to handle larger volumes...              DNA sequencing drives our bioinformatics needs       Fig. 1: QC'ing data at Addgene is like...been crucial for us to keep pace with our current need to analyze nearly 300 sequences each week. Our internal... challenge: automating the bioinformatics steps needed to transform the raw sequencing data into a complete...Seqera’s mission and our own, as both address the need to accelerate science using different approaches...

...spent on details that do not yet need to be decided. Figure out what you need to accomplish to get to the ... meeting at all. When no discussion is needed and you just need to give everyone a heads up (for example...everyone’s time by considering the level of meeting you need. Project Meetings: You do not have to include every...workload, updates on current projects, and offer feedback. The employee should ask for input on projects...the manager know if any additional resources are needed. All-Company Meetings: As your organization grows...whole thing together. How long do you actually need? Just like liquids, meetings tend to fill the shape...of their container. Consider whether you actually need a full hour to achieve the purpose of the meeting...

...single most important skill that scientists need to succeed in their work. While it's not always recognized...scientist needs to be able to communicate with their colleagues and peers. Scientists also need to be able...substitute for feedback from others.  Seek out members of your target audience and ask for their feedback before...will have in the world is the value that you can succeed in communicating. Even the most rigorous, insightful... interactions, ask yourself how every piece of feedback you receive generalizes.  If someone suggests ...   To address that 'why' question effectively, you need to make your audience the focus of every sentence... write and step you take.  Why does the audience need to know this? How will they react to it - what will...

...testing new products or methods, using metrics and feedback, taking a look at what works and what doesn’t,...can guide viewers through. I know cameras, but we needed someone else. Design: Jennifer thankfully volunteered.... It’s wonderful and simple to use, but when you need to juggle filming and performing on camera, it’s...No scripting made this a lot easier and I didn’t need to be there directing. Everyone’s schedules were...While this works well for social media videos, we need as much detail as possible when showing lab methods...  Design: The previous cycle made me realize we needed to focus on ways to shoot safely at a distance ...out of reach for Addgene.  So, onto research! I needed to design a system that would allow me to set up...

...that need to be resolved. If there are any forms that need to be signed, or manuscripts that need to be...their ideas confidently while also considering the needs of other people. Some students and postdoctoral .... It is in your supervisor’s interest that you succeed, and if you cannot resolve a problem on your own...reach out for help so you can get the support you need. 2) Prepare an agenda for every meeting Supervisors...brainstorm about solutions that will meet both of your needs. 6) Put important agreements in writing One of ...meeting with an email that summarizes what you have agreed upon and your action items. This will give your... what you have discussed and add suggestions if needed. 7) Always follow through on your end of the deal...

...kits are guaranteed to yield endotoxin-free DNA. Once you know the type of DNA prep you need, you can ... right for you. What type of plasmid prep do you need? As we covered in a previous post on plasmid preps...downstream application will dictate how much DNA you need to recover and the level of quality required. Some...Figure 1 illustrates the three grades of DNA you may need based on their relative quality and quantity.   ... and up). But perhaps more importantly, the DNA needs to be free of contaminants, such as bacterial endotoxins...as their input (Table 1). When you feel like you need just a bit higher DNA yield, but not enough to go...increasing your input to increase your output. But, exceeding a kit’s capacity can actually reduce your ultimate...