... Basically, if you can devise an experimental assay that works with flies, you can utilize the resources... Seahorse can be employed to look at oxygen consumption. Drosophila are also used in cancer research (... I’ll provide a brief introduction as to how you can engineer new transgenic flies to study your favorite...others use attP sequence docking sites so that you can control exactly in which locus your transgene ends...1): Random insertion has the advantage that you can get varying expression levels of your transgene in...for genetic recombination so that the transgenes can be inherited on the same chromosome. Targeted Insertion...which chromosome your transgene has integrated. This can be accomplished with a few matings to balancer chromosome...

...everyone can access data and drafts of manuscripts and grant applications in real-time so they can provide...Science is a global endeavor, and that can often mean that scientists who live across the country or ... these accelerate the progress of research, they can also present hurdles for the team of researchers ...disciplines or doing different tasks. But these challenges can be exaggerated when communicating across time zones... that helped make my experience run smoothly and can help other teams successfully overcome these challenges...every month is long enough that some experiments can get done and data analyzed between each meeting, ...conducting the experiments to the PIs. Remote meetings can be tricky, and it takes extra effort to make sure...

...online meeting can take some of that pressure off. This question and answer process can also help you ...campus even if you can’t visit. By talking with people at the institution, you can get a better sense ...the in-person portion got canceled, taking advantage of virtual meetings can still be a great opportunity... flood of interview requests and visitation days can be, at times, very overwhelming. As someone who applied...technologies that helped us navigate a global pandemic can also be the thing that provides a new perspective... applications opening. With virtual options, you can narrow down your list of schools by meeting with ... idea of that advisor’s research interests which can be helpful in creating your application to that program...

...). While some mutations can remain stable as homozygous alleles, others cannot. Geneticists need a method...happens. Is it an evolutionarily conserved gene? Can you find an ortholog in the Drosophila genome? Continue...Continue reading and I’ll show you how Drosophila can be used to push your research in new and exciting...of seeing a phenotype since additional genes that can do the same function are less likely to exist). If...system), and the third post will describe how you can make your own mutant flies. Find Drosophila Resources...~9 days after setting up the genetic cross). You can tell when pupae are ready to hatch the next day because...their bodies will be very light in coloration and can look very plump. On the side of their abdomen will...

...Both of these can be increased by concentrating a viral stock. Different titering methods can reveal differences...virus solution can be obtained. 4. Enhancing virus-cell contact A variety of reagents can increase transduction..., you can check if virus is being generated by your transfected packaging cells. This check can be performed...An estimated 320,000 viruses can infect mammals. Even more abundant are the Earth’s estimated 1031 bacteriophages... viruses are everywhere and doing everything, it can be annoying when we try to use them in an experiment...which are factors in the technical challenges they can present. Many researchers find themselves plagued...titers (which is unavoidable for some vectors) you can concentrate your virus by ultracentrifugation of ...

...mutagenesis is a highly versatile technique that can be used to introduce specific nucleotide substitutions...or deletions) in a tailored manner. The approach can be used in conventional cloning (to introduce or ...contains the template, site-directed mutagenesis can be used to mutate the PAM sequence (an NGG sequence...Site Directed Mutagenesis In brief, point-mutations can be introduced to plasmids using primers (with the...plasmid containing the mutation). Similar results can be obtained with PCR screening using the primers ...repetitive sequences, but if present, a minor extension can usually ensure that the 3’-base(s) do not form secondary...larger constructs, but plasmids as large as ~6 kb can be amplified fairly easily by simply following the...

... This step can involve multiple smaller steps that all help to ensure that antibodies can access their... that are not exclusive to IF, but they can have a significant impact on the outcome of your IF experiment...bind to their targets, most commonly proteins. You can then use fluorescent microscopy to look for those...ICC are essentially just more general terms that can also refer to assays using non-fluorescent visualization...polyclonal, which means that multiple secondary molecules can bind to a single primary, resulting in signal amplification...of possible primary antibodies that exist and you can begin to see why generating fluorophore-conjugated...made in different species on a single sample, you can then use the corresponding secondary antibodies conjugated...

...biotinylated proteins can be captured on streptavidin affinity beads or matrices. The bait protein can even target...2018). BioID and BioID2 can take hours to label proteins and APEX2, although it can rapidly label proteins...the mgSrtA variant can be displayed on the surface of a cell of interest, where it can label interacting...affinity of biotin for avidin, proximity labelling can capture weak/transient protein interactions as well...used for both soluble and insoluble proteins, which can be a challenge for classic antibody based affinity...methods.  BioID: a promiscuous biotin ligase that can efficiently label neighboring proteins The first ...proteins. Thus to capture protein complexes, scientists can simply express a bait protein fused to BirA* in cells...

...when appropriate, and can be saved at any point throughout the process, so you can work through it at your... antibody, the Data Hub can help you decide if an antibody is a good candidate for your experiment. If...data on which might work in your application. It can be quite difficult to decide on which antibody to...’s Antibody Data Hub Addgene’s Antibody Data Hub can help with this decision. In the open-access Data ...researchers interested in the Addgene antibody collection can look at user-deposited data detailing how an antibody...of the Addgene Data Hub.   Antibody data reports can be searched and sorted by gene/target, name, application... conditions in which the data was generated, you can scroll down the page to the materials and methods...

...one pod can pick up 96 tips for easy sample transfer between 96 well plates. The second pod can pick up...simple steps you can take to automate processes in your own lab. Some things your lab can do to get into...to do to find said robot and to find out how you can get one. As a starting point, I would recommend conferences...is it’s customizability. It has a large deck that can house many different types of devices. As long as...’s easy to pipette samples in and out of it, you can set up a procedure for it using the Biomek FxP. For... is pretty simple. By clicking a few buttons you can quickly tell the Biomek FxP how much sample to pick... making it easy to spread out your samples. This can be useful if you’re transferring samples from 96 ...

... They can result in lively discussions with both panelists and audience fully engaged. They can also be..., but the side can work. Only put out enough seats for the expected attendees (you can have extras ready...and program organizers can help make sure the event stays on schedule. People can only sit so long. I recommend...audience playing Candy Crush or reading email). My advice is always "don't do it unless you can do it well"... food or a fantastic view (these are nice if you can afford them). I am talking about making sure the ...interesting stories, riffs or opinions that each panelist can contribute. The moderator should identify the funny...panelists of the stories or topics you know they can speak about. There should be some spontaneity, but...

...things you can do to prepare. Adapt to WFH while parenting Keep your kids on a schedule so you can rely ...that you can adapt to your kids' needs. Plan ahead. Think about and set up activities that kids can do more... week to flatten the curve, but working remotely can be more difficult if your work is based in the lab...outdoors is so huge, these are activities that you can easily do while staying away from others.  Meditate...assist in labs that are testing for COVID-19. You can also give back by supporting local businesses (ex...until now.  Network. There’s a lot of networking you can do online. See if a scientist or someone you’re connected...writing a full paper, that’s great. Otherwise, you can begin working on these drafts and for a few model...

...interspaced with linkers that can fold into a secondary structure that can bind specific proteins via aptamer...CRISPRi system can be repurposed to include modular RNA scaffolds. These RNA scaffolds can recruit various... to include modular synthetic RNA scaffolds that can recruit various transcriptional regulators (Zalatan...expression, multiple scRNA transcriptional programs can be regulated with dCas9 as a master ON/OFF switch...module, alternative sets of transcriptional programs can be layered with distinct programs controlled by each...expression at the targeted site. Multiple scaffold RNAs can be designed to allow simultaneous, multidirectional...tunable control of gene expression. This strategy can be applied to effectively change metabolic flux. ...

...free avidin can be added to your sample. Any free biotin-binding sites on this complex can bind to your...biotin is added to a protein or macromolecule. Biotin can be added enzymatically or chemically. Enzymatic biotinylation... in the egg white. Avidin is a glycoprotein that can bind four molecules of biotin.   In the late ...glycoprotein that consists of four identical subunits that can each bind one molecule of biotin. Biotin and avidin...added to. Biotin also has an exposed side chain that can be easily manipulated to make reactive species without...polypeptide is biotinylated (more on this below) it can be incubated or co-expressed with avidin or a structurally...the 1960s, strepavidin. The Biotin-avidin complex can then be used and/or manipulated for a scientist’s...

... streptavidin, both of which can bind up to four biotin molecules and can themselves be linked to a labeling... sodium azide, which can interfere with conjugation reactions. If needed, you can often remove the incompatible...spectrum, so they are great for multiplexing and can be used in a variety of assay types. Be sure to check...antibody conjugates in an earlier post, where you can find some additional tips for working with these ... which is light produced by a chemical reaction, can be detected using specialized imaging instruments...mass cytometry (Bendall, et al., 2012). And, you can also conjugate antibodies directly to beads for purification... linked to your antibody. This chemical coupling can be achieved through several different mechanisms....

...collection can be used for in vivo phenotypic screening of small-molecule libraries and can be combined...monogenic diseases. Additionally, retroviral vectors can regulate gene expression by delivering shRNA or miRNA...and the 2nd and 3rd Generation Lentiviral Systems can be found in our Lentiviral guide. Popular packaging...RNA-mediated gene silencing mechanism. Scientists can express siRNAs to direct the cellular RNAi machinery...Cre-recombinase. The Cre-lox system is a technology that can be used to induce site-specific recombination events...gene of interest pLenti-puro (Plasmid #39481) You can clone your gene of interest into this tetracycline...lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors (empty backbones...

...enough that their bulk can be a barrier at times. Smaller, slimmer affinity reagents can have deeper penetration..., meaning that each scFv can target a different antigen; binding of each can be either simultaneous or...kD but can have weak signals due to their monovalent nature. They are stable up to 80°C and can refold...readily available for a large number of targets, and can require specific conjugates. And, of course, once...often used in clinical applications. Fab fragments can be created in lab in situations where a suitable ...orientation of the two fragments. However, rigidity can be introduced through mutation, allowing diabodies...repeats, have high specificity and binding affinity, can be produced in E. coli, and are both soluble and ...

...for clear information on costs. Can you suggest some coaches? Yes I can. Please note that Addgene does ... many scientists.  Sometimes it can be helpful having a coach that can offer extra support for issues ...Professional Career Coaching can be an excellent tactic for scientists making a career shift or who wish...wish to improve their current job situation. What can a coach do that is different from an adviser, boss...accessible. Sometimes only a few hours of coaching time can make an enormous difference so don’t assume that ...offices, postdoc offices or professional organizations can provide referrals. There are bad coaches out there... a good one that is a style match for your needs can take some research.  It helps to spend some time ...

...that can be done.  Note that each type of transfer requires different equipment. Wet transfers can be done... and can help amplify a weak signal. However, they have high cross-reactivity, vary significantly from...light-insensitive. They can detect femtogram levels of protein. These enzymes cannot be used for multiplexing...? What membrane? What should you block with? It can be so overwhelming that you might just stick with...monoclonal Both polyclonal and monoclonal antibodies can work well for a western blot. Polyclonal antibodies...batch-to-batch variation. Confirmation of sequence can be done from relatively cheap plasmid sequencing....of different rAbs that target the same protein — can be used to reproduce the benefits of polyclonal antibodies...

...procedure can be used to incorporate each element into the final construct. This strategy can be used to... each other, a third module cannot be placed between them, but rather, can only be added to the 5’ or ...synthetic biology for example, high-throughput cloning can be used to combine the functions of different genetic...reporter constructs. These fluorescent reporters can be used to detect structure and protein localization...sequences, and more. By combining these components, one can generate constructs to visualize cell cycle dynamics...directional, meaning that (in the previous example) one can control whether the third module is added to the ...References Golden Gate Type II restriction enzymes Can assemble up to 10 modules in a single step. Through...