...Hands-off super busy types, who never have time to mentor you, and Micromanagers, who question every detail...the needs of other people. Some students and postdoctoral fellows are hesitant about expressing their ... 5) Listen to your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor...Look at the problem from their viewpoint, and brainstorm about solutions that will meet both of your needs...other supervisors might be nice people, but poor mentors (super-agreeable friendly types, indecisives, and...graduate students since 2009. Dora is also the creator of the “Finish Your Thesis” online course.  To ... Read Joanne Kamen's Post on What Makes a Good Mentor Resources on the Addgene Wesbite Find plasmids...

...particular surface receptor are stained with a fluorescent antibody against the receptor and analyzed on ...fluorochrome is removed from all other detectors except its dedicated detector. To ensure proper compensation ...directions. The degree of scatter is captured by detectors and provides useful information about the cell...emits a fluorescent signal that is captured by detectors. The strength of the emitted photons varies from...to further separate the B cell population into progenitor B cells, immature B cells, plasma cells or others...flow cytometry experiment depends on a variety of factors including sample preparation, staining procedures...

...Gateway pDEST, and pBAD vectors for example. Protein expression from each of these vectors is controlled by ...which expression system to use depends on several factors: The protein you are trying to express How much... interest is inserted into a plasmid expression vector that is then transformed into a bacterial cell....cellular debris. There are several popular DNA vectors that can be used to produce large amounts of protein...resulting in different levels of expression from each vector; lower expression may be required if your protein...protein is toxic to E. coli. Of all the vectors, pET, under the control of the T7 lac promoter and induced...

...with a Cas3 effector, type III with a Cas10 effector, and type IV with an unknown effector. Type I has...repertoire of CRISPR has brought many other Cas effectors to light. One lesser-known Cas is waiting for...genomic alterations on a scale that other Cas effectors can’t, while maintaining relatively high efficiency... family, which means it requires multiple Cas effectors to carry out genomic editing. Specifically, these...still be a challenge. A multitasking Cas Most Cas effectors have one job — Cas see DNA, Cas chop DNA. Cas3...

...Zhang Lab - Multiplexed Overexpression of Regulatory Factors (MORF) Library 3,548 250 - 10,000 bp,...more difficult to transform.    Figure 1: Factors to consider when creating a pooled library amplification...Created with BioRender.com.   The most important factor to consider is your desired library coverage, since... type of cells you are using, as well as other factors, such as your lab’s normal transformation protocol...protocol, the size of the cuvettes your lab’s electroporator uses, etc. The DNA amount can be anywhere from...inserts of different sizes (such as a transcription factor overexpression library) or is prone to recombination...

... by guest bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Golden gate assembly...restriction enzyme specificities at each step. Factors such as enzyme efficiency, stability, and buffer...four base overhangs after cutting. The destination vector receives the assembly after similar Type IIS restriction... inserts. After ligation of the insert into the vector, the BsaI site is no longer present, allowing multiple...the original insert religates into its original vector, it will subsequently be re-digested and "eligible...our guest bloggers Becky Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Becky Kucera is...assembly, New England Biolabs, Inc.         Eric Cantor is a molecular biologist and a Development Group...

...in 2004, we were the plasmid repository. In 2016, we added viral vectors, and in 2022 we added ready-to-use...ready-to-use antibodies. Now, we think of ourselves as a repository for DNA-based materials, including our popular... kits and libraries. This DNA-based materials repository contains 139,868 reagents from 5,821 labs in ...international community, we've partnered with distributors in China, Korea, Japan, India, and Brazil.  ... system for scientists using materials in our repository, from our customer and science support to our...

...amazing stories to share. I spend a lot of time telling scientists about Addgene’s repository, and explaining...student, post-doctoral fellow, lab manager, research technician, or Principal Investigator. How-to guides...for genome editing? Know a professor with a great story to tell? Let us know by emailing us at [email protected] on Twitter and Facebook. We’ll track down the story, get the inside scoop, and keep you informed.   ...

...significant holidays. Figure 4: Plasmid request trajectories suggest an exponential decay demand model with... scientists or labs placing orders.  Our model factors the likelihood in our Poisson process into two ... multiplies the likelihood by the corresponding factor 𝜙z if t is during event z. The "events" we considered...each region using stochastic gradient descent in Pytorch, which gave us the maximum likelihood estimates...chemistry and chemical biology.       David R. Liu is Director of the Merkin Institute and Vice-Chair of the ...University; and Howard Hughes Medical Institute Investigator. Liu’s research integrates chemistry and evolution...four examples of technologies pioneered in his laboratory. Learn more here. Additional resources on the...

...m a father of three, a scientist and science communicator, and I strive to be a volunteer and contribute...scientist. When our elementary school needed a new coordinator for the annual Math & Science Night, I took the...contacting our school district’s science curriculum coordinator with minimal luck, but eventually the 8th grade...for a scientist. I think that establishing this history helped demonstrate that a) I was a real person ...great feedback from teachers and other science communicators.  What can CRISPR do? I spent more time on...which demonstrated to the students that scientific history was very much in the making. One of the exciting... in an exciting, tumultuous time in scientific history. Whatever your reasons, let us know how it goes...

.... (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation of Fusion Constructs ... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...expression system Choosing an appropriate expression vector is the first step in generating transgenic Arabidopsis...from the Tsuyoshi Nakagawa lab are binary gateway vectors and can be used for promoter swapping and generating... ‘A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology’, Frontiers in Plant...

... Bank (DSHB), directed by David R. Soll, is a repository for monoclonal antibodies, distributing over ...antibodies to be used as a powerful reagent in the laboratory.   The DSHB focuses on monoclonal antibodies,...include those directed against human transcription factors such as PAX7, PAX6, AP-2 alpha and ISLET-1. In ...against new cancer biomarkers. We believe that repositories, such as Addgene and the DSHB, play an important... to ensuring affordable access to materials, repositories provide standardized quality-control and technical...to Our Guest Blogger!   David Soll has been the Director of the Developmental Studies Hybridoma Bank (DSHB...

...Mammalian Vectors By Marcy Patrick Will You Be My Mentor? Finding and Asking for Mentoring Support ...through the lens of Addgene’s blog and plasmid repository. Our blog is just over a year old, and it has...more and more scientists offering to share their stories – about their research, their job hunts, and their... Genes By Marcy Patrick Plasmids 101: Yeast Vectors By Marcy Patrick CRISPR-Cas9 FAQs Answered! ...

...proteins (FP) have also been important in mapping, monitoring, and manipulating neuronal circuits. Epitope ... is therefore difficult to use epitope tags to monitor single molecules - something researchers often ...tags, low FP expression may be insufficient for monitoring a weakly expressed POI. Spaghetti Monster - A... CA3 pyramidal neurons which are known to be notoriously difficult to label. It has also been shown that...technologies such as array tomography, super-resolution STORM imaging, and electron microscopy. Sample preparation...standard proteins tags like GFP. Specifically for STORM, the high intensity signal provided by smFPs can...Spaghetti Monster probes at Addgene Browse all AAV vectors Find ready-to-use AAV preps ...

...Rogers is a postdoctoral researcher in Patrick Müller’s group at the FriedrichMiescher Laboratory of the Max... post was contributed by Katherine Rogers, a postdoctoral researcher at the Friedrich Miescher Lab of ... response to an experimental treatment can be monitored. This can reveal previously unknown functions ...splicing. (RNAi, in contrast, engages existing gene regulatory mechanisms to block activity and does not work...phiC31. The popular UAS/Gal4 and Cre/Lox gene regulatory systems have also been successfully implemented..."Single-cell reconstruction of developmental trajectories during zebrafish embryogenesis." Science 360.6392...

...reinforcement are ICSS and place preference.   Historically, ICSS was done by implanting an electrode and...successful inhibition of hippocampal neurons. Initial monitoring Second, this may sound rudimentary, but before...exciting—and unanticipated—road of discovery. Monitoring behavioral output The appropriate behavioral ...shock that has become associated with a visual or auditory cue. The hippocampus has long been known to be...versatility and standardization of genetic tools in the laboratory mouse, most optogenetics is performed in this...University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne Kreek at the....    References 1. Stuber, Garret D., et al. "Excitatory transmission from the amygdala to nucleus accumbens...

...Our plasmid repository succeeds because of our amazing community. The trust of our depositors to allow us...focused on our mission – documenting your plasmids, storing them, providing quality control and customer support...more scientists outside of the US now use the repository – about 50% of plasmids are shipped to labs outside...manage our website, and continue to improve the repository. We love being able to provide a useful (and ... us to help store and distribute their plasmids. The partnerships and collaborations with institutions...

...Of course, all of you toiling away in laboratories this holiday season want the work you are doing to...been to publish in journals with a high “impact factor,” a measure that dates back to 1975 and is based... online access on scientific impact Partly, the story has to do with online accessibility. It is now about... recently reported that the most cited work in history is a 1951 description of an assay to determine ...of scientific publications. Ventures like Impact Story seek to find ways to capture scholarly work that... impact factor...

...of new tools came out including the single base editors, Casilio, CombiGEM, and a variety of pooled libraries...top requested plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9 from the Zhang Lab.... TLDR: This is a versatile mammalian expression vector that you can use to express both SpCas9 and your...resistance gene as separate peptides. PX459 This is the vector backbone. This particular backbone can be used ...Overall, this is a versatile mammalian expression vector that you can use to express both SpCas9 and your...mammalian cell lines. Alternative forms of this vector contain GFP-tagged and un-tagged versions of SpCas9...

...Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia Research...tribulations of the process, I’ve provided guidance and mentoring to several people that found my insights helpful...It helps to come from the lab of a well-known investigator. If you only have middle author papers or no... potential collaborations, critiques of your postdoctoral research, etc. The second question is whether...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum, optimization...