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TypeBlog PostPublishedOct. 20, 2020, 1:15 p.m....record yourself. You can watch yourself to critique and figure out what to improve. You can share it with others...charge before others do”. Candidates who have “started something” which they can share on their resumes ...There are many different kinds of grants that you can apply for. If you are in a lab, your PI is likely...to work-from-home and just now realizing that we can’t bide our time and wait for it to be over to work...roles but for almost every position, we look for candidates that have demonstrated initiative. In fact, hear...have a definite advantage in our process. We see candidates who have done the following types of things: ...relationships when you are more desperate or needy so you can’t wait until you imminently need a job. Start now....
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Cultivating Community Science at BosLab
TypeBlog PostPublishedNov. 2, 2017, 1:35 p.m....what appears to be a funky old garage, but looks can be deceiving. Step into this building and you will...volunteers. Transforming citizens into scientists Anyone can become a member of BosLab and previous laboratory...environment, and openly publish protocols so that anyone can engineer yeast to do similar things. The focus is... To learn more about what we’re up to or how you can help BosLab, visit our website and follow us on twitter... -
Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids
TypeBlog PostPublishedNov. 20, 2018, 8:17 p.m....this can lead to wasted time or retractions. Using a catalog number for identifying a reagent can be helpful...specific Addgene ID # listed in a publication, you can quickly pull up the associated plasmid page on the...87155 ; RRID:Addgene_87155).” So when citing, you can copy and paste the text attributing the PI who generated... -
Hot Plasmids: FRET-Based Biosensors
TypeBlog PostPublishedMay 6, 2014, 1:07 p.m....calcium sensors, which are available at Addgene, can be used for ratiometric in vivo imaging. Griesbeck...on" in the presence of calcium. This color change can be observed under a microscope using commercially... beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans, and zebrafish...and his colleagues took to optimize sensitivities can now be applied to dozens of other published FRET-... -
Advice for Scientists Starting a Lab
TypeBlog PostPublishedApril 29, 2014, 1:43 p.m....recommends to seek out advice from seasoned PIs who can offer support. “It is an exciting and emotional time... not rush when growing your lab. “One bad choice can be counter-productive." He also emphasizes the point... at about 65,000 messages and dealing with email can be half of my day sometimes.” He points out that ... to companies.” Take home message Starting a lab can be a challenging process. Each PI emphasizes the ... team so getting everyone talking to each other cannot be undervalued.” Connie Cepko points out the limited... -
Science Communication for Everyone
TypeBlog PostPublishedApril 16, 2019, 1:05 p.m....process of starting a forum where non-scientists can directly approach scientists and doctors with their...visual representation of science to our audience. How can researchers work with you? We welcome researchers...contributing to The Addictive Brain. A contribution can either be an article, a short post, a video, or a... -
Making Career Development an Integral Part of the Science Training System
TypeBlog PostPublishedFeb. 25, 2021, 2:15 p.m....a student or postdoc ends up in a lab where they can learn the plus skills needed to succeed on the career...disseminating career training approaches in a way that can be implemented widely in the community. Talk alone...scheduled for March 4th from 2:30-4:00 pm ET. You can learn more about the report's recommendations and... -
A Conference By Postdocs For Postdocs: Future of Research
TypeBlog PostPublishedSept. 2, 2014, 3:05 p.m.... we have to change to ensure that all scientists can have successful careers doing science in some form...the upcoming "Future of Research" conference. You can register for the conference here. The Future of Research...programs or peer mentoring groups. But for now, you can click here to register and join your postdoc colleagues... -
6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results
TypeBlog PostPublishedJune 26, 2014, 2:23 p.m....DNA sequencing troubleshooting and analysis 1. You can use any of the following programs to view your .ab1...all inclusive service some companies offer), you can purify the product either via the Sodium Acetate/...strongly recommend using a silica spin column. They can be pricey, but well worth it. 6. Edit your DNA ... -
Twenty Years of Addgene Sharing: CRISPR
TypeBlog PostPublishedApril 18, 2024, 1:15 p.m....tools. Distributing and depositing In Figure 1, you can see cumulative numbers of CRISPR plasmid distribution...categorized by function, from 2014 to 2023. As you can see, nucleases, the original CRISPR tool, remain ...the most popular with our requestors. However, we can again see quick adaptation and steady use of all ...categorized by “age” or year they were deposited, you can see that from 2013 and on, popular CRISPR plasmids...ordering the plasmids is the easy part; using them can be a bit more challenging. Addgene’s free educational...other countries, from Japan to Australia, are significant depositors and requestors. CRISPR tools are developed... -
AAV Vector Quality Control: Going the Extra Mile with NGS
TypeBlog PostPublishedSept. 12, 2017, 1:44 p.m....cell genome can be packaged inside the vectors during production. These impurities cannot be removed during...capabilities and Seqwell’s plexWell technology, we can now perform NGS at Addgene to sequence all the DNA.... In most cases, the source of the contamination can be identified, and we will discard any AAV prep we...developed custom open-access Python scripts that can perform a deeper analysis of the data. Our serotype...Next-generation sequencing (NGS) is a powerful tool that can be used to identify DNA contaminants in AAV preparations... -
A History of Genome Engineering in Popular Culture
TypeBlog PostPublishedFeb. 25, 2020, 2:15 p.m....were first designed in 2005. The DNA-binding domain can be engineered to target specific DNA sequences while...then induces a double strand break. These breaks can then be repaired by homologous recombination with...triplet and when several ZFNs are strung together, it can read many base pairs simultaneously to increase specificity...geneticist named Zobrist who designs a virus that can potentially kill one-third of the human beings (just...rely on the cellular DNA repair machinery, which can lead to high frequency of indel formation in the ...called pegRNA (prime editing RNA), the prime editor can install all possible point mutations, delete short... understanding of the adenine base editors which can help accelerate the engineering of novel base editor... -
A Control for All Seasons
TypeBlog PostPublishedOct. 5, 2023, 1:15 p.m....critical control in biological experiments. Replicates can be technical or biological and are included to improve...such as bovine serum albumin. Secondary antibodies can also react with endogenous antibodies naturally present...in mouse tissue. Such species-on-species staining can be partially addressed with stringent staining protocols...way to reduce, if not eliminate, the noise. This can be done either by modifying the blocking conditions...autofluorescence, also interferes with data interpretation and can be addressed with a label control. In a label control...allows you to detect dead or dying cells so that you can base your analysis on living cells only (for example...results, the signal from your target needs to be significantly higher than the background noise (Waters 2009... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog PostPublishedSept. 28, 2023, 1:15 p.m.... to specific subcellular regions, laser ablation can damage other cellular structures, and genetic mutations...Restricting the effect to a specific time or location can be critical when, say, the gene is essential during...with dozens of family members and many functions it can be difficult to unravel their specific roles. Zebrafish...using directed evolution (Tei et al., 2023). PLD can hydrolyze phosphatidylcholine (PC), the predominant... membrane lipid, to phosphatidic acid (PA) or it can swap phosphatidylchonine’s choline head group for...to plasma membrane or lysosomes and its activity can be monitored by phosphatidic acid probe GFP-PASS.... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog PostPublishedJune 3, 2021, 1:15 p.m..... ScFvs can also be fused to cellular toxins, radioisotopes, cytokines, and enzymes for cancer, autoimmune...scFv that binds cancer-related antigens, binding of the scFv part of the receptor can activate T cells...are the smallest unit of an antibody molecule that can bind antigen. While they may seem unfamiliar, scFvs...both VL-linker-VH and VH-linker-VL configurations can generate functional scFvs; however, some individual...scFvs are easier and cheaper to make because they can be expressed in bacteria, while mAbs generally require... Power., 2019). Their rapid clearance from blood can be a drawback for therapeutic applications where ...cells to kill cancer cells. scFv’s small size also allows for their delivery by viral vectors like AAV, which... -
Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?
TypeBlog PostPublishedMay 10, 2016, 2:30 p.m....infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral therapies such that ...from infected cells. Read on to learn how CRISPR can take on HIV-1 in vitro, and what obstacles must be...healthy individuals. Antiretroviral therapy (ART) can virtually eliminate plasma HIV-1, improving life ...promoter-driven GFP expression. In cell lines that can produce HIV-1 toxic proteins, CRISPR/Cas9 SAM caused...our reach. Similar studies have shown that CRISPR can be used to combat other viral infections, notably...learned in pursuing one technological application can benefit many others and accelerate the pace of research...inflammation, and negative metabolic effects of ART. Why can’t the body mount an immune response to destroy these... -
Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons
TypeBlog PostPublishedApril 24, 2015, 2:39 p.m....Without Cre recombination, RFP is expressed. Cre can mediate one of two deletions to allow YFP or M-CFP...using 2 loxP variants (only identical loxP sites can mediate recombination.) The deletions are mutually...deletion and inversion. The Brainbow-2.1 construct can express one of four colors (n-GFP, RFP, YFP or M-... two color possibilities. The remaining cassette can invert as long as Cre remains expressed; once Cre...includes a mutant YFP that does not fluoresce, but can be detected via immunostaining. This feature facilitates...placed under different promoters, Brainbow and Flpbow can be used to label distinct cell populations. To decrease...three recombinations, and subsequent recombinations cannot occur. While the Brainbow-1 system employs Cre-... -
Addgene’s Viral Service - Why Virus? Why Now?
TypeBlog PostPublishedMarch 23, 2017, 2:30 p.m.... approaches everything; we asked ourselves, “How can we do this in a way that most helps scientists and...about virus production protocols and quality control can be found on Addgene’s Viral Production webpage). ...partnerships to expand the service and figure out what we can afford to do and when. Addgene’s Legal Team acquired...Requested Lentivirus and AAV of 2016 Learn How You Can Use the Viral Service in Your Research Using Virus... -
DAP arrays for multiplex gene editing
TypeBlog PostPublishedJuly 7, 2022, 1:15 p.m.... promoters, has limited the number of gRNAs that can be packaged into viral vectors for potential in vivo...the Gao Lab demonstrated that the tRNA-gRNA array can drive the expression of itself, without using any...gene editing with DAP array Use DAP arrays can enable efficient multiplex gene editing, such as ...different tRNAs with optimized 5’ leader sequences can be used together in DAP array. To construct DAP array...their initial testing. It outperformed other tRNA candidates (either from human or plant sources) in both ... -
New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor
TypeBlog PostPublishedApril 7, 2022, 1:15 p.m....a variety of roles in synaptic communication and can trigger other forms of neuronal signaling and regulation... The yellow (v82) and lime green (v857) versions can be summarized quickly as: iGluSnFR3 v82: increased...being deposited at Addgene. Other plasmids and pAAVs can be ordered directly from Addgene. The Podgorski Lab...provided a brief breakdown of the promoter systems you can use to express the iGluSnFR3 variants: pRSET: Bacterial...