...been ordered more than 20 times; red, more than 50; and blue, more than 100 times! When you search for plasmids... plasmids which have been ordered more than 100 times).     I guess it's cool to be popular, but how ... number (if known), antibiotic resistance, and optimal growth temperature are listed, as well as the name...

... CRISPR use, you will want to invest some time in optimizing and testing the efficiency of CRISPR delivery...Ribonucleoproteins (RNPs) Read Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User) Resources on Addgene....

... teaching responsibilities left him with limited time for one-on-one mentoring. His solution? Bringing...particularly amenable to in-class labs. Provost ultimately settled on a two-semester model, in which students...Maximizes Reach Many new professors are wary of the time and effort and/or perceived professional risk involved...try implementing one. It's likely only a matter of time before a CURE trainee goes on to win a Nobel!   ...

...grant professionals have similar stories about a time when their passion for an organization or a program...area of expertise. Whether you’re seeking a full-time job or want to be an independent grant consultant...and grant management.  Your niche will change over time. Going into the grants profession with a concept... will save you and the organization(s) you serve time and resources. 3. Get a mentor, find your people...

...they were ten years ago, and it does take a bit of time to craft one. We still think they're useful, so ...applying to the specific company or lab. Take some time to research the lab or company via their papers ... to start from a standard version, but take some time to make it fit the job you’re applying for. The ...professional. Always review grammar and spelling several times and ask someone else read your cover letter (as ...

...However, the two different approaches allow for optimized strategies for differing amounts of target proteins...following the incubation step. The wash buffer has an optimal pH and ionic strength that breaks weak interactions...antibody intact. The wash step is repeated several times to ensure non-specific proteins are completely removed...isotype control antibody. Alternative applications Oftentimes, an IP-appropriate antibody simply isn’t available...

...during cell division simply does not allow for enough time for correct partitioning to occur (Diaz et al., ... low copy plasmid is more likely to be lost over time.  Is plasmid incompatibility going to affect my ...clarity, a wet lab transformation experiment will ultimately inform whether your plasmids are compatible or...beginning to use plasmid incompatibility to combat antimicrobial resistance. Using novel plasmids based partly...

...efficiencies (Wang et al., 2013). Although you can optimize your primers for each individual sample, this ...ensure accurate and reliable results, we have optimized our assay in a few ways:  Absolute quantification...titrating each lot of AAV in vivo to determine the optimal dosage. At Addgene, we don’t reserve lots, but ...quantification by droplet digital PCR versus analog real-time PCR. Nat Methods. 2013 Oct;10(10):1003-5. PubMed...

...software development resources into it. At the same time, this project gives Snapgene great exposure to the... part of my work. However, this is not the first time I’ve had a role in organizing collaborations. During...companies and then convince the contacts to give me some time to talk or to at least read my emails. Then pitching...Personally, I find this to be the most challenging and time-intensive part of relationship building. However...

...gotten to know what a plasmid is in general, it's time to take a look at some of its parts. Elements such...promoters are constitutively active and on all the time while others are more carefully controlled. Regulated...might act only in certain tissues or at certain times in development or there may be ways to turn them...promoters available to scientists. Plasmids are oftentimes put to therapeutic uses, and in those cases it's...

...vectors can be produced easily, without the need for time-consuming purification processes, and are powerful...integrate into the host cell’s genome. However, sometimes inte-great-ion isn’t so great. In this post, we...expression of gene products (Wu et al., 2004). Over time, the expression diminishes as the episomes are lost...express a protein or nucleic acid for only a limited time, IDLVs are ideal. As the episomes are lost during...

...Addgene! Since the development of BioID and APEX, optimized versions of BioID and APEX have been developed...promiscuous biotin ligase The Roux Lab in 2016 optimized BioID to create BioID2, a smaller and improved...hypothesized that the low sensitivity was due to suboptimal folding or stability. Thus they conducted multiple... significantly greater than miniTurbo at earlier time points but miniTurbo exhibits less labeling prior...

...hold a doctorate degree, are currently working full-time, and have received a research grant in the last ... down when dropped will not make it happen. In a time when a top official can be quoted as saying “I’m...primary contributor' to global warming The New York Times: With Trump in Charge, Climate Change References...Technology: Crossing The Imaginary Line The New York Times: A Scientists’ March on Washington Is a Bad Idea...

...cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes...technique, and is less prone to off-target mutations. Ultimately, successful allelic exchange requires two sequential...mutants, but it frequently requires considerable optimization to ensure strong counter-selection. Anecdotally...magenta. This is particularly valuable when first optimizing conjugation in strains that don’t conjugate that...

...Each cell remains the same color for its entire lifetime and daughter cells retain the same color, allowing...allowing for the fate mapping of cell populations over time. The ability to track single cell dynamics at the...to create foci of proliferating cells. Tetbow optimizes Brainbow for current technologies Tetbow (Sakaguchi...that incorporates chemical tags instead of XFP to optimize it for tissue clearing, a method that makes tissues...

...and this is likely a rate limiting step, but optimization of the exon-splitting site sequence can enhance...and acceptor sequences were tested and with the optimal sequences, the group achieved ~80% of muscle cells...level of transgene expression, and may need to be optimized for each unique application (Trapani et al., 2015...resources on the Addgene blog: Find tips for a first time AAV user Learn about how AAV is used in retinal ...

... and Lichtman estimate that Brainbow’s colorful labeling has decreased the mapping time for a given section... were limited due to low resolution or were very time-intensive, necessitating the development of a new...is truly fitting for this colorful technology. Optimizing the system: Brainbow-3 Although Brainbow provided...

...behaviors, but little is known about the order or timing of these factors. Recombinase-based state machines...reaction can only be performed on one register at a time. If a particular application makes it difficult ...machine design space is highly degenerate and often times there are multiple register designs that may be ...? We tested a two-input (ATc and Ara) RSM over a time series of increasing input exposure lengths and ...

...with linear DNA as repair templates which reduces time to generate edits to only four days (Paix et al.... rapid. This stringent selection allowed me to optimize a range of editing parameters: oligo repair templates...Farboud et al., 2019). The Meyer lab also determined optimal PAM orientation for editing using two Cas9 targets...Resources Learn about gRNA design and gRNA optimization for your CRISPR experiment ...

...2015). Meanwhile, Sanger sequencing is laborious, time consuming and cannot be applied to heterogenous ...multiplexing) Requires large cell population, senstive to time since cell fixing HTGTS Not reported Cell-based...sequencing techniques, please see “Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity” by Tycko...Vic E. Myer, and Patrick D. Hsu. "Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity." Molecular...

...inconclusive results, erroneous conclusions, and wasted time. In fact, many have called out antibodies as major...and so this approach may not be best if you are ultimately planning to use your antibody to detect endogenous... proceed with confidence.  Share your results Ultimately, like all of science, validation experiments ...could work great in another context! So the next time you test an antibody (perhaps one from Addgene!)...

...as confusing as reading Shakespeare for the first time. The discovery and application of CRISPR-Cas9 kicked...bacterial and archaeal immune systems. During this time, scientists discovered many related immune mechanisms... only recently that researchers began building optimal plasmids to harness the power of Class 1 systems... your goal, experience, hypothesis, and even the time and money available. The breadth and diversity of...

...synthesized and degraded within the cell, so at any given time, an FP-chimeric protein may be at any one of many...quenching”). The photoswitch can occur multiple times by activating with two distinct wavelengths of light...possible excitation lasers) it is also important to optimize your activation protocol for your particular set...heavily on the same fluorophore being imaged multiple times in order to reconstruct an image. The best PA-FPs...

...distributed over 800,000 plasmids since 2004. In that time, our lab has gotten a lot experience in processes...tip #1: Glove trick Putting on lab gloves can sometimes be annoying. To make it easier, try this quick...a single tube and simply flick the tube several times. This will help ensure ensure equal distribution...

...is recognizing the PAM on DNA, the CTD is also sometimes called the PAM-interacting (PI) domain. The guide...al., 2013). As mentioned above, however, Cas9 sometimes tolerates mismatches in the next 8-10 bases after...and the DNA sequence is recognized, it’s finally time to cut! The HNH domain, which cuts the target DNA...Shima, T., Nureki, O., & Uemura, S. (2018). Real‐time observation of flexible domain movements in CRISPR–Cas9...

...contributed by Kusumika (Kushi) Mukherjee. The ultimate goal in the field of regenerative medicine is ...iPSCs into the cell type of choice. This step is sometimes also referred to as “directed differentiation....39]. In 2000, Ferber et al. showed for the first time that mouse liver cells could be transdifferentiated...Partial rearrangement from one cell type to another Time and resources More (due to additional pluripotency...

...Thoughts on his time in Boston and Outro We'd like to thank Dr. Kwapata for taking the time to talk to us...across the globe. Listen here! Podcast Breakdown (Time: Topic) 0:00-1:00 : Intro 1:01-3:48 : Kingdom Kwapata's...

...here at Addgene HQ in Massachusetts that means it’s time for near-freezing temperatures, our first of the...decorating contest! Our #DeckTheLab contest is a time for scientists worldwide to show off their decorating...little more festive this holiday season! It is that time of the year again: miniature #Christmas tree goes...

...graduate student life. Senior graduate students can sometimes be elusive or have a certain mystique that makes...Listen to the interview here! Podcast Breakdown (Time : Topic) 0:00 - 0:28 : Niroshi and Ben describe ...We'd like to thank Niroshi and Ben for taking the time to sit down and talk to us and wish them the best...

...origin. The type of cloning approach you choose ultimately will influence the quantity of DNA that is required...viability, reduces transfection efficiency, and ultimately affects the quality and outcomes of experiments...plasmid DNA, so that you can decide what will be the optimal type of plasmid preparation in terms of quantity...