...avoid bleeding as much as possible. However, when you need to label multiple markers, some bleeding is often...cytometry is commonly referred to as bleeding. When two fluorophores bleed into each other, it means that the...different target proteins in your flow analysis, you'll need to use fluorophores that attach to the target. When... When you design a flow experiment, you'll need to pick a fluorophore for each target and ensure the fluorophores...properly label many markers with any fluorophore. Bleeding Before we start building your first panel, we'... a protein and FITC an organic molecule), their bleed is so strong that these spectra almost overlap. ...As eGFP emits fluorescence by itself and does not need to be bound through a separate fluorophore, we have...

...Horticulutre Biotechnology, he did his PhD in Plant Breeding, Genetics, and Biotechnology at Michigan State...award from nonprofit and Addgene collaborator, Seeding Labs, for lab equipment that will help accelerate...about Dr. Kwapata’s current work and thoughts on the need for more scientific collaboration across the globe...:41 : Kingdom on the difficulties surrouding and need for biotech research in Malawi 11:41-14:53 : Kingdom's...challenges in Malawi, and more 23:45-28:07 : How Seeding Labs is helping advance Kingdom's research 28:08... Kwapata for taking the time to talk to us and Seeding Labs for connecting us with him. Learn more about...about our recent collaboration with Seeding Labs here. Special thanks to our sound engineer/producer, Eddy...

...competitive CV You don't necessarily need a Nature/Cell/Science paper. You do need multiple first/corresponding...solicited for feedback), possibly the Dean, and even some people in your field that still need to understand...members will NOT be in your area. Therefore, documents need to minimize jargon. Equally importantly, you really...me that their search committee is different and needs to be told all of the nuances of the research in...Bottom line is to appreciate that your audience needs to understand your proposal to give you proper consideration...your research because you waded too deep into the weeds. Check out Joanne Kamens's "10 Steps to a Perfect...idea in a swamp of unnecessary details. You DO NOT need to present a detailed review of your field or the...

...Luna, Joan Reede, Alberto Roca, and Jessica Tytell made it was obvious that more data is needed for us to...outcomes. Some of the more specific contours of what we need to know in order to even assess the current state...the professional skills and research independence needed to pursue his or her chosen career path,” but individual...companies and organizations (including Addgene), that need people who are data oriented, scientifically minded...graduate students and postdocs. Trainees however, need to be made more aware of the fact that these positions...highlights just this point. As part of this, trainees need to be able to network and interact more with people...well as young PIs and all gave advice on how to succeed in publishing in the current academic environment...

...hear you saying “We know this already!” But, you do need to be able to research and write your application...fresh eyes and correct it (and, believe me, you will need to correct it). It’s easy for reviewers to spot ...department) to read over your application and give you feedback. They’ve most likely been through the process ... your area are looking for. Remember though, you need to leave them time to read it, give it back to you...Funding agencies want value for their money so you need to make it clear that you will be doing your research...you have been working for the last few years, you need to be very clear that you are staying because it...the world for your research to rapidly progress. Need Help Building Your Scientific Network? Check out...

...-international focus. Candidates in need of an industry H-1B need to be aware of the April 1 deadline ...necessary and no job offer is needed for this category. To qualify, you need to satisfy 3 out of the 10 ... their work in the US have the added burden of needing sponsorship. If you are in this position, you should... green cards that are well-tailored to meet the needs of researchers. The categories are as follows: ...organizations can be a good choice for individuals needing sponsorship. Academic H1B The academic H-1B allows...your case – the invitation to be a peer reviewer needs to be addressed to your own personal email directly... not allow for individuals to self-petition. You need the support of your institute to do this one. Universities...

...: expert No-needle techniques The term “injection” is most often associated with needles, but it may ...surprise you that they don’t always require needles. If needles make you cringe or your experiment will allow...in mice. Image created with biorender.com.   Needle techniques Intravenous Intravenous (IV) injections...actually requires more technical skill to place the needle in the correct vein without collapsing it. Retro-orbital... (IT) injections are probably the most advanced needle injection technique. Intrathecal injections are... allow you to avoid poking the mice, these “no needles” techniques may come in handy. These techniques...technique involves administering medication through a feeding tube with a bulb on the end, which prevents accidental...

...list. Need feedback on their work but prefer to be approached as peers rather than subordinates Need mental... is hard and most people have some learning they need to do before they can be good at it. Like any other...work harder and contribute more. Assuming basic needs are being met (i.e. enough pay and benefits to live...to work mostly at home. Having a high degree of freedom and diversity built into their jobs Chance to do... to identify the work style of each team member) Need challenging work, opportunities to pursue and problems...people, not just scientists) Are self-directed, but need precise leadership and support from their superiors...comfortable with imposed deadlines (there’s that need for flexibility, sometimes taken to an extreme by...

...aliquoted to reduce the number of freeze-thaw cycles needed when using the virus. A small aliquot is used to...hundreds to millions of distinct plasmids, all of which need to be represented in the same proportion after amplification...library can drastically impact what conditions it needs to properly amplify.  Amplification protocols therefore...protocol, or have constructed your own library, you’ll need to develop your own protocol that amplifies without...Verification Once your library has been amplified, you'll need to verify your library by next-generation sequencing...If you’re ordering a pre-made library, you don’t need to worry about amplifying and producing lentivirus...libraries aren’t quite plug-and-play: you’ll still need to titer and optimize before you can transduce your...

...addition to all of the documents you submit, you will need to present a "chalk talk." However, few, if any,...search committee member cannot either. Therefore, you need to be able to describe experimental approaches in... if the aim is successful. Note that aims do not need to be grant aims. Each aim could be the basis of...Once you have developed your research vision, you need to package it as a roughly 30 minute chalk talk....interesting. For example, “there is a significant need for healthy livers for transplants. Growth of new...What will you have accomplished in 5-10 years? You need to be able to describe what success looks like. ...Describe the question, the bare minimum of background needed to understand the problem and goal, the approach...

... degrading them. You’ll also need to know the wavelength of laser needed to excite them - and it’s a good...experiment - there are a number of outside factors that need to be considered as well. In this post, we’ll touch...antibody. This information is important, and you’ll need to track it throughout your experiments.  Now that...and biological structures, some of which you may need in your experiment.  If you are planning to conjugate...light sensitive, so antibodies conjugated to them need to be stored either in a dark, opaque vial or wrapped...experiment is easy, but what about those times you need more than one?   Multiplex assays Multiplex assays...each with a different signaling molecule, that all need to be detected in the same assay. This is one of...

...valuable feedback.  Listen. Think about the questions you’ve received, read the comments, seek feedback from...amount of mental thought and effort your audience needs to understand what you’re communicating. While I...striving to obtain?” If the answer is yes, you will need to spell out that connection for your audience. ...blank spaces or to gloss over some aspects when needed. The trick is to communicate your key points well...students know.   Ask the learners themselves for feedback in a variety of ways, like having them restate...about how I was presenting the information. Maybe I needed to remove a distraction, such as pausing or turning...something was intuitive that actually wasn't. Maybe I needed to employ a visual aid instead of describing something...

...experimental system used, so you’ll need to know what expression pattern you need, and the factors in your system...cells you’re targeting; (3) how much expression is needed to read your experimental output; and (4) the consequences... translate to in vivo efficacy. You’ll therefore need to test potential AAV vectors in your specific experimental... AAV2 (Watakabe et al., 2015) (Figure 1). You’ll need to consider if such properties could help, or harm...affected by the cell and/or tissue type, so you’ll need to test for this in your system if you’re measuring...some neurons can transiently express TH - so you’ll need to check for this in your system and then test each...doxycycline (Favre et al., 2002). They are useful when you need to control the timing of expression in your experiment...

...Of course, you’ll need to align all these images once taken. To do so, you will need to include a marker...beautiful images? Multiplex IF may be just what you need! How does it work? There are several ways to achieve... in one step. As with standard IF, you will need to start with a fixed sample. Depending on the sample...sample protocol.)  Simultaneous IF Depending on your needs, your sample, and targets, you might decide to do...) is more flexible and efficient, since you only need a few fluorescent secondaries on hand in the lab...species, isotype specificity, and dye meets your needs and budget, or you can make your own. For example... polyclonal) Goat anti-Rabbit IgG   If you need more signal, another strategy is to use a biotinylated...

...often offer higher allelic specificity, due to the need for the target:probe/primer bonds to be more stable...ll want to predict this value using BLAST, you’ll need to experimentally verify once you’ve received the...Validation Once designed, your primer and probe set needs to be validated and your reaction optimized. You...treatments may affect the assay. It’s unusual to need to adjust amounts of reagents in your chemistry,...the bench As a highly sensitive technique, qPCR needs a steady pair of hands to produce reliable and reproducible...rapid-cycle machine such as a LightCycler, you’ll likely need to re-optimize protocols for that machine. (Teo,...from understanding how the analyses work - you do need to make sure the assumptions in your dataset match...

..., you should get feedback from those who attended. You may even seek, or get feedback from those who didn...current or enjoy that discussion. Luckily, one needn't worry if they choose to move away from academic...data, software repositories, or other information needed by someone to critically discuss the paper.  Before...meeting, now what? After the first meeting solicit feedback. To ensure that the journal club is useful for...attend. For example, we got a good bit of early feedback from those who wanted, but were unable to attend...find a time that accommodates more people. The feedback could be solicited via multiple avenues including...there are a few things you can point to. Does the feedback you receive include the sense that people learned...

...probably don’t need it to express your protein of interest. However, you do still need a species-compatible...In fast-growing populations of cells, the vector needs to replicate alongside its host, or it will be lost... DNA replication begins, so a bacterial plasmid needs a compatible ori that will be recognized by its ...the host cell’s genome, so no special sequence is needed to retain the vector as cells divide. In transient...transient transfection may be sufficient for your needs even without a compatible replication site. Regardless... contexts. For a vector to persist in... You need... Bacteria a compatible ori Yeast an ...plasmid is delivered to your target cells, you still need to select for the cells that contain it. Antibiotic...

...the steps needed for TOPO cloning: 1. Create Your PCR Product: Design standard primers (no need to add unique...organized and prepare everything you need for the next step before proceeding. Pre-warming your antibiotic-containing...results in the production of blue colonies that do not need to be picked and sequenced for potential positive...Do not add 5’ phosphates to your PCR primers; you need that free hydroxyl group! You may want to include...all unpaired 3’ ends in your PCR product. If you need to decrease error rate, please use one of these ...room temperature, it is not recommended that you exceed the 5 minute time limit (lower transformation efficiencies...reported with longer incubation); however, you may need to incubate for 20-30 minutes if your PCR product...

...accept feedback and adapt their mentoring styles to the mentee’s personality and professional needs at any... science as well as the developmental needs and resources needed to pursue them, increased their network...undergraduate and graduate studies. Students also need more access to reputable, organized networks connecting...international experts and students. In response to such needs, the International Mentorship Program (IMP) USA-... successful example. For evidence of this, you needn’t look any further than testimonials from the students...Program work? IMP Pilot Program 2015-2015 Scholars need strong proactive mentors who have relevant expertise...group of individuals with technical and career path needs, and came from the universities mentioned above....

...allowing repair to take place before DNA replication. Indeed, they observed that 41/42 embryos were non-mosaic... at the mutation site, indicating that repair proceeded through NHEJ rather than HDR. Figure 1:...: Homology-directed repair in targeted zygotes proceeds from the maternal allele rather than the supplied... characterization studies, the edited embryos proceeded to the blastocyst stage at similar rates to untreated...surrounding CRISPR-based therapy is the level of testing needed to define whether off-target editing or mosaicism...not possible with IVF, as most of the cells are needed for proper development. Ma et al.’s experiments...experiments showed a 3% mosaic rate, and this rate would need to be much, much lower before clinical applications...