...ask for an amplification protocol with your deposit. This is because repeated amplifications of pooled ...for further downstream applications. As individual libraries can vary significantly, it’s important that... an amplification protocol developed for each individual pooled library; generalized amplification protocols...smaller libraries, chemical transformations can also be used. Efficiency for chemical transformations is...that Addgene distributes with a chemical transformation amplification protocol is the Chong Zhang E. coli...Plasmids Variable Region Size DNA per Amplification Amplification Type Link to Protocol CRISPR Knockout... this blog, we’ll discuss how to develop an amplification protocol for your pooled library deposit. When...

... was founded in early 2018 with the goal of communicating science to non-scientists. Chinmaya Sadangi,...questions about The Addictive Brain and science communication for general audiences. How does The Addictive...Addictive Brain address science communication? At The Addictive Brain, we collaborate with scientists from around...disorders are frequently overshadowed by other medical conditions, so these topics have become a cornerstone...sci-art.  What are your top tips for science communication? It is important to identify your audience ...on the Addgene blog Find all of our science communication posts Sign up to receive our science career ...guide Learn about different careers in science communication Resources on Addgene.org Learn how Addgene...

...original publication and ensure that it includes validation data relevant to your application — and double-check... use a mechanical method before your enzyme lysis. For cell cultures, enzyme lysis is typically sufficient...for your experiment. But if you understand the technical aspects of a western blot, you'll be able to understand... so let's dive in!      Figure 1: The technical decisions to make when designing a western blot...However, they have high cross-reactivity, vary significantly from lot-to-lot, and have to be re-validated...multiplexing is challenging and limited. Direct westerns typically rely on primary antibodies conjugated to fluorescent... antibody or loading sample, which may not be practical. Conjugated primary antibodies can be more difficult...

...researchers involved. Communication is a major challenge when researchers are communicating across disciplines...closely replicates how you’d interact with team members in person, and improves overall communication.  Maintain...Furthermore, this casual communication reflects the type of informal communication you’d likely have with...But these challenges can be exaggerated when communicating across time zones, digitally, and asynchronously...to keep everyone on the same page is to make communication a non-negotiable part of your collaboration.... Maintain informal communication In between recurring meetings, you should share interesting results or..., this may happen over email or using rapid communication tools such as Slack. Share data and documents...

... DNA purification without a kit as well as one way to reuse silica columns from DNA purification kits....that silica spin column, stop to consider some ways to purify DNA without a kit. DNA purification kits.... Kit-less DNA purification methods avoid many of the drawbacks of kits and typically follow the same...plasmid purification and DNA gel extraction method described in Kim and Morrison is nearly identical to that... DNA to silica columns, it’s bound to glass syringe filters. Normally DNA does not bind silica or glass...et al., 2018. Regeneration of silica columns for DNA purification Starting material: PCR product or...method for regenerating silica columns that come with PCR product purification or DNA gel extraction kits...

...Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously...create specific edits in DNA, there has also been significant work on constructing Cas9 transcriptional activators...reassemble when exposed to specific UV wavelengths or chemicals (Polstein and Gersbach, 2015; Zetsche et al., ... Tuttle and Alex Chavez! Marcelle Tuttle is a medical student at Tufts University and former researcher...researcher at the Wyss Institute for Biologically Inspired Engineering.     Alejandro (Alex) Chavez is a member... (2014) A Protein-Tagging System for Signal Amplification in Gene Expression and Fluorescence Imaging....

... A lot of companies and organizations selling biological reagents have a ton of data on the catalog page...originally described in a peer-reviewed scientific publication. We love science, but we don’t have the resources...it’s own tongue-in-cheek website.  If you are skeptical about using Google for serious science questions...companies have real scientists answering their technical support questions (besides Addgene, I know both...be fairly fast, getting the full answer to a complicated question may take some time. If the customer ...message Answering customer service questions is a delicate balance of knowledge, patience, and diplomacy,... is answerable, particularly when it comes to technical questions. BUT, I do believe that scientists who...

...previous nosy question! The Application Process Broadly speaking, the PhD application process tends to go something...steps in the application process and usually occurs in the few months leading up to applications opening. ...program. Most applications will recommend that you name two professors in your application that you intend...Virtual) Campus Visits After receiving applications, universities typically invite a select group of students...to continue the application process through a campus visit, a weekend where applications can get to know...of reasons (logistical, financial, etc…) now have options to fully engage in the application process. And...strengthen your application and help you decide if a program is a good fit for you. The application process looks...

...the origin of replication/replicon. The replicon is comprised of the origin of replication (ori) and all...   Types of origins of replication There are lots of origins of replication out there so, for simplicity...gene for replication (reference). **F1 is a phage-derived ori that allows for the replication and packaging...control elements. The ori is the place where DNA replication begins, enabling a plasmid to reproduce itself...are generally different from the those used to replicate the host's chromosomal DNA, but they still rely...readily at lower temperatures and allows the replication machinery room to come in and get busy making...derivatives), pSC101, R6K, and 15A. Not all origins of replication are created equal. Some will produce many plasmid...

...plasmids rely on their host cell's replication machinery in order to replicate—but not always to the same extent...described in our previous Origin of Replication post, DNA replication is initiated at the ori and may or...consider the replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...review how E. coli normally replicate their chromosomal DNA. Chromosomal replication in bacteria is carefully...entire round of replication. Initiation is the primary control point of DNA replication in bacteria, so..., it recruits helicases to continue unwinding the strands. This creates two replication forks that move...chromosomal DNA ensure that the helicases are released from the DNA once the replication forks reach each other...

...names for technical support roles. Examples are Technical Service Representative, Technical Support Specialist...these departments from technical support. Technical support careers in brief Technical support careers are...troubleshoot problems at the bench. Specifically, roles in technical support make excellent use of the ...worked as a technical support specialist for 4 years. Read on for insights from my time in technical support...products successfully. In technical support you may have to answer technical enquiries such as: “Can I... areas of expertise in technical support positions. When applying for technical support positions, you...familiarize yourself with technical support. Even having personal experience with technical support through the...

...re more interested in selling a product than communicating your ideas with people or discussing science...I do a lot of public speaking where I enjoy communicating face to face with the audience. I post on LinkedIn...own expertise and your resources for science communication opportunities like an AMA. One of the best things...recommend taking every opportunity you get to speak publically about your work (you can find my tips for a perfect... science communication...

...the chemical factor, of a naturally occurring voltage indicator. To give an example, a genetically encoded... Gong Y. Enhanced genetically encoded voltage indicators advance their applications in neuroscience. Curr...and sensing electrical potential. Thankfully, there is a tool to measure these electrical events! Voltage...Voltage indicators are voltmeters for cells! In this blog we will review how these indicators work, their...their general applications, and why viruses are essential to their use as biological tools. Why Use Voltage...Voltage Indicators? Electrical signals are integral to many facets of biology. For example, many heart issues...studying voltage within a cell are genetically encoded voltage indicators (GEVIs) and chemigenetic sensors...

...and includes biochemical properties, previously established uses, and intended applications. Within CasPEDIA...CasPEDIA, there is a classification system, CasID, to help facilitate direct comparison of different enzymes...multiplex ability Experimental guidelines and applications Protein structure and properties Links to ...you want to read literature on the less common applications of an enzyme for your non-traditional experiment...al. (2023). CasPEDIA Database: A Functional Classification System for Class 2 CRISPR-Cas Enzymes [Preprint...I., Iranzo, J., et al. (2019). Evolutionary classification of CRISPR–Cas systems: a burst of class 2 and...

...after application of NE (10 μM) and corresponding pixel-wise ΔF/F0 heatmaps. White insets indicate surface...developing and depositing two new norepinephrine indicators: nLightG and nLightR (Kagiampaki et al., 2023...surface expression of the indicators over white dashed lines. Scale bars, 10 μm (HEK293T), 20 μm (neurons...trialed in two-photon in vivo experiments. nLight indicators are more sensitive to and selective for norepinephrine...feedback in response to norepinephrine release, and clinically-used drugs targeting these receptors cannot be...They can, however, be studied with the nLight indicators. Speed One of the major advantages of this next...nLightR, both generating a measurable signal after electrical stimulation with a single pulse, while nLightR...

... Clinical applications Viruses in vaccines helped pave the way for their use in other clinical applications...etc.). When you’ve decided on an application, our Viruses as Biological Tools blog can help you learn about...vector from basic science research all the way to clinical gene therapy. But how can you use these amazing...s unpack the myriad number of emerging viral applications and give you some recommendations for putting...putting viruses to work. Viral applications Viruses work by infecting a host cell (the target of viral infection...and delivering a genetic payload. This cargo is typically their own genome, but this step can be engineered...expressed. It’s important to note that for all the applications we discuss, there is more than one type of virus...

...– The basics Statistical analysis is what determines if there is statistical significance among data sets...values – numerical risk allowance in a statistical test. Distribution - spread of data; typically, normal...asterisk of significance? You’ve already done the heavy lifting, so don’t let the statistical analysis of... sets. Significance is a numerical value which quantifies the likelihood that a result is due to chance...are statistically different. They are classic parametric tests and one of the simplest statistical tests...analysis on a five-sample data set yields statistical significance, you won’t know if only two of those samples...if the dependent variables worked synergistically, antagonistically, were additive, etc.). ANCOVA and...

...xkcd.com Science communication should be fun and engaging for the communicator as well as the audience...many ways options available to today's science communications: podcasts, blogging, videos, lectures, exhibits...in what people think of as "the best way" to communicate their science, meaning that they will heavily...heavily evangelize for the mode of communication they connect best to. And that's really natural - if it works... text or use Facebook. However you choose to communicate your science, some people will love it and others...audience, so pick a mode of communication that you enjoy and that plays at least slightly to your strengths...outside of the box. There are so many ways to communicate and I wholeheartedly encourage experimentation...

...extension is repeated 25-35 times to exponentially replicate the target DNA of interest. This entire reaction...have been developed for different scientific applications. These PCR methods all use the same basic PCR...not as quantitative as other methods of PCR. Theoretically scientists should be able to determine the quantity...the final cycles which would slow or stop PCR amplification. Other times, the PCR reaction may not be 100%...quantified by qPCR. qPCR is used in a variety of applications including gene expression profiling, studying...concentration, unlike qPCR where results can vary across replicates. ddPCR can be used to quantify DNA sequences ...droplet contains the material required for PCR amplification. Following the PCR reactions each droplet is...

...protein purification. For a more thorough introduction to pCri, please see the original publication cited...expression and purification: a comprehensive review of affinity tags and microbial applications." Biotechnology... Protein purification can be one of the most stressful lab activities. Working with proteins requires...interest can save a lot of time later. Protein purification specialists recommend doing some research, notably...Gomis-Rüth, aims to take the guesswork out of protein purification through simple and efficient multiplexing. pCri...sites should one need to remove the tag post-purification. A sample of pCri constructs: pCri-1a, -4a ...4a and -9a. 6xHis tags enable affinity purification using a Ni-NTA resin. MBP improves protein solubility...