...lab, or maybe even training on a new technique with a coworker. Being a new trainee (even if you’re an...an oldbe trainee!) can be pretty nerve-wracking. So how can you be the best trainee possible? It’s not...your best self to the training experience.  The ins and outs of being a trainee There is a lot of advice...true for training in the lab! If you don’t do proper preparation and pay attention during training, then ...questions to your training! All of these things will help you get more out of the in-person training component...need to double-check a detail! Training follow up After your training session (ideally in the next few... the information in your training. Bench mentor-mentee relationships Training can be a very fulfilling...

... which is ease of transformation. There are many motivations to generate transgenic Arabidopsis, from ...from studying transcriptional and translational dynamics of genes and proteins in living plants, to complementing... dip transformation method. The general steps for this method include: Cloning and transforming a plasmid...integrates transfer DNA (tDNA) into the genomes of the plants it attacks Growing the transformed agrobacterium...obtain optimum transformation efficiency. It can take 6-8 weeks from the date of transformation to having ...2013). Preparing plants for transformation Whatever plants you are transforming into will be your T0 generation... ready to transform when they begin flowering. It is important to perform the transformation not long ...

...generally are not transmitted to adjacent neurons via synapses (i.e., transmitted transsynaptically). This is ...Converting a transsynaptic neuronal tracer into a monosynaptic (first order) neuronal tracer For RABV to... connections. While this transsynaptic spread is useful for neuronal tracing, it has two practical limitations...retrograde transmission (Etessami et al., 2000, Wickersham et al., 2007). Thus, RABV transsynaptic spread ...operates. References 1. Callaway EM. Transneuronal Circuit Tracing with Neurotropic Viruses. Curr Opin ...Retrograde neuronal tracing with a deletion-mutant rabies virus. Retrograde neuronal tracing with a deletion-mutant...neurons are connected. Viral Vectors as Neuronal Tracers Due to their ability to efficiently enter cells...

...of a frustrating situation. Asking for opinions from a neutral third party can be a useful strategy to ...researchers feel they lack formal training in a variety of transferable skills. At Addgene, we've set out...You can easily work on this transferable skill without any formal training or coursework. First, you have...signing contracts when working for a business development team, negotiation is a key transferable skill ... by highlighting that researchers learn many transferable skills while working in the lab and by offering...where you might need some help. Today in our transferable skills guide: Negotiation. As a grad student...business suits discussing multimillion dollar contracts behind closed doors. Fig. 1: The mythical...

...triple T’s of biology are transformations, transfections, and transductions! In this blog we will discuss...intricacies of transductions, we've got you covered there too. What is a transduction?  Transduction is the ...Molecular basis of transduction Transductions utilize the ‘normal’ mechanism of viral infiltration into a cell... experiment. Using transduction vs. another method Transfection and transduction seem to achieve similar...picky when it comes to transfection/transduction methods. Some ‘difficult to transfect’ cells lines are amenable... process of transferring foreign nucleic acid into a cell by a viral agent. Transductions are similar ... then sterile filtered to be extra sure nothing untoward gets transduced (like a rogue HEK-293 cell). ...

...requires further study. Traditional neuronal tracing methods Traditional tracing compounds either produce...retrograde tracing take advantage of existing transport pathways in neurons. Anterograde transport is typically..., and enzymes for transmitter synthesis. Retrograde transport is used for transport of endocytosed material... facilitate transport: retrograde transport relies on dynein, while anterograde transport relies on kinesin...mechanisms of trans-synaptic trafficking are not fully understood and may be a form of transcytosis where the...cell after intracellular transport without actually causing infection or leading to transgene expression... neuronal tracing experiments, brain tissue is injected with compounds that intracellularly spread away...

...immediately prior to their transduction has been shown to increase AAV transduction efficiency by approximately... 3. Transduction efficiency depends on viral titer Both of these can be increased by concentrating a viral...unavoidable for some vectors) you can concentrate your virus by ultracentrifugation of the viral stock, followed...before proceeding to ultracentrifugation to avoid contaminating your transduction with the packaging cells... by ultracentrifugation (75,000 - 225,000 g for 1.5 – 4 hours at 4 °C). After ultracentrifugation, virus...PBS.  Alternatively, to concentrate a virus solution without ultracentrifugation, you can reduce the volume...immediately following transfection. In other words, when exchanging the medium after transfecting cells, replace...

...Blog Transferable Skills Guide: Problem-Solving Transferable Skills Guide: Teamwork Transferable Skills...researchers feel they lack formal training in a variety of transferable skills. At Addgene we're helping...creativity is absolutely a transferable skill developed during PhD training. Creativity is required to... to tackle it. It’s easy to get distracted by an email or sidetracked by a colleague. To innovate, you...both helping researchers recognize their MANY transferable skills learned while working in the lab and ...Today, we’re highlighting an oft-underlooked transferable skill: creativity. I’ve heard quite a few scientists...any scientist. In this blog, we’ll delve into strategies you can use to foster innovation. If you identify...

...researchers feel that they lack formal training in a variety of transferable skills. At Addgene we've set out...Chiara, the tech transfer expert who understands the somewhat complicated Material Transfer Agreement process... highlighting that researchers do learn MANY transferable skills while working in the lab and by offering... skills of many of my colleagues, I can also orchestrate the formation of high-functioning-but-temporary...technical customer service. Team members are highly trained, and though there are necessary redundancies in...once famously said “I think this is the most extraordinary collection of talent, of human knowledge, that...own experience: Leadership Few things are more frustrating than being on a leaderless team. Sometimes the...

... to avoid contamination, take extra care when performing RNA extraction. RNA is inherently not as stable...expensive ultracentrifuge.  Figure 1: An outline of the different steps in RNA extraction. Researchers...your extractions- if you need really clean RNA, it’s important to make sure that when extracting, to only...stable as DNA- it is single stranded and its ribose groups are susceptible to hydrolysis and heat degradation... They will degrade in a -20°C freezer.   RNA extraction methods evolved into a simple protocol still ...without a kit. However, that isn’t the case for RNA extraction and purification. There is one simple method ... loaded onto cesium-chloride gradients and ultracentrifuged for up to 20 hours to separate the intact ...

..., exo-AAV2 in mouse retina after intravitreal injection to transduce bipolar cells and photoreceptors ...22314290. PubMed Central PMCID: PMC3345986. Meliani, Amine, et al. "Enhanced liver gene transfer and evasion...PubMed PMID: 29296848. PubMed Central PMCID: PMC5728288. Schiller, Lara Timantra, et al. "Enhanced Production... vectors. They found that they have a higher transduction efficiency, i.e. the same number of AAV particles...particles is able to infect more cells (as demonstrated in different cellular environments such as retina...also found that exo-AAVs were more resistant to neutralizing anti-AAV antibodies. This is particularly useful...vivo where endogenous anti-AAV antibodies often neutralize the therapeutic efficacy of AAV vectors. How ...

...apply to programs, and apply for travel grants. Good luck and happy travels! Resources Resources on the ...remote collaborations Transferable Skills Guide: External collaborations Transferable Skills Guide: Teamwork...current institute. A common reason is hands-on training in a technique that only a few labs have expertise...this expertise will provide you with guidance and training so that you yourself can become an expert and ...for another reason. Visiting would allow you to train and acquire the data you need on the new instrumentation...new collaborations and fresh perspectives. For trainee level researchers, it can expose them to a new ... Identify reagents you will need, any required training, and a priority list of experiments you hope to...

...resources on the Addgene blog Transferable Skills Guide: Time Management Transferable Skills Guide: Conflict ...of being a scientist and an incredibly useful transferable skill. Being able to stand up in front of an... audience and convey your ideas clearly and strategically is powerful in almost any career. It allows ... applies: what can go wrong, will go wrong. The trash removal service slams dumpsters outside the open... turn your slide deck into a chalk talk. Your translator doesn’t know the term “diving board” so you end...messing up?–and use the three tips above to make a strategy that can help. Even something as simple as telling...remember to breathe!)   Public speaking as a transferable skill The worst thing about doing public speaking...

...Posts in the Transferable Skills Guide Developing Transferable Skills During Science Training Resources...researchers feel that they lack formal training in a variety of transferable skills. At Addgene we've set out... highlighting that researchers do learn MANY transferable skills while working in the lab and by offering...where you might need some help. Today in our transferable skills guide: Collaborating with others outside...of my position at Addgene I am now involved in Strategic Partnership Management, a very interesting and...being clear about what was expected, and keeping track of speakers. During your graduate or postdoctoral...perfect platform for practicing initiative and for training in teamwork and collaboration management. For ...

... The higher the titer, the better. AAV transport and transgene expression It’s clear that we need high... the issue of low titer by concentrating your virus. See our Concentrating AAV Protocol (Step 10) for ...PubMed Central PMCID: PMC2574934. 3. Zingg, Brian, et al. "AAV-mediated anterograde transsynaptic tagging...contact with the cell is to increase the AAV concentration. In this flowchart, you can see the dwindling...be required. For example, retrograde AAV gets transported up the cell’s axon. While the mechanism for this...yet fully understood, we can imagine that this transport is probably not 100% efficient. So the few AAV...particles that have entered the cell now have to travel up the axon, through the cell, and potentially ...

...researchers feel that they lack formal training in a variety of transferable skills. At Addgene we've set out... highlighting that researchers do learn MANY transferable skills while working in the lab and by offering...where you might need some help. Today in our transferable skills guide: managing a team. You’ve just been...about management skills during your scientific training. I never thought about management skills before...grad school. Both were hard-working scientists-in-training who got the job done, but I struggled with what... he graduated. Years later I attended a formal training for new managers and I had a “Eureka” moment while...around the fact that new team members need to be trained, which may seem like a waste of your time at first...

...friendships. I put “neutral” in quotations here because I would not have considered myself neutral in any of their... blog Management for ScientistsTransferable Skills Guide: LeadershipTransferable Skills Guide: Teamwork...stopped talking to each other if I was not there to arbitrate for them. They were different people with some... them would acknowledge later that without my “neutral” presence to alleviate those differences, they ...the right time. Like many of the posts in our Transferable Skills series, this post is not meant to be ... that you could strengthen. The right set of transferable skills, coupled with your technical skills, ...The source of those conflicts can frequently be traced back to one of a few trouble spots:  Communication...

...researchers feel they lack formal training in a variety of transferable skills. At Addgene, we've set out...evaluate potential strategies to resolve the problem.  Step Three: Decide on a strategy and implement it... highlighting that researchers do learn MANY transferable skills while working in the lab and by offering...where you might need some help. Today in our transferable skills guide: Problem-Solving. I don’t know ...potential sources of error and then decide on a strategy to fix the issue. I was problem solving!  You ... careers, I always felt that descriptions of transferable skills seemed fuzzy or ill-defined. In this ...it.  Step Four: Assess how effective your strategy was. If you didn’t quite fix everything or revealed...

...express a transposase or reverse transcriptase so they can transpose. Both class I and II transposons can be...eukaryotic genomes.  Components of a transposon system Transposon: The transposon is the DNA sequence that moves... encodes the proteins the transposon needs for transposition. Transposition protein: This is the protein...themselves as RNA transcripts and then rely on a reverse transcriptase enzyme to reverse transcribe back into ...reverse transcriptase and integrase enzymes which are required for LTR retrotransposon transposition. Non-LTR...reverse transcriptase activities.    Figure 1: Overview of retrotransposon transposition. Retrotransposons...2021. To mobilize a transposon, first the transposase enzyme binds the transposon’s long terminal repeats...

...easier to track translation in two different ways: by monitoring the first round of translation or by tracking...of relative reporter translation. Single molecule continuous translation tracking TRICK allows you to ...various stimuli on global translation, but a visual technique to study translation remained elusive. Two ...destination co- or post-translationally. TRICK: Visualizing the first round of translation While ribosome profiling...in the fluorescent signal: untranslated RNAs appear yellow, and translated RNAs appear red. To enable ...Using translation inhibitors cycloheximide and puromycin, Halstead et al. verified that translation is necessary...to see the first round of translation, but it doesn’t allow continued tracking of a given mRNA because ...