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TypeBlog PostPublishedJan. 14, 2016, 3:30 p.m....plasmid are the backbone, inserted gene/functional mutations, antibiotic resistance, restriction sites used...most sequence assembly errors occur), deletions, mutations, in-frame protein fusions, and multiple cloning...cases, we deal with multiple inserts or internal mutations that require more than two sequencing reactions...
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Career Coaching for Scientists: Why and Where Do I Find One?
TypeBlog PostPublishedJan. 29, 2015, 1:25 p.m....A professional coach will provide an initial consultation meeting or call for free. Take advantage of...affordable for you. There is also a group The Dissertation Coach based in Illinois. A few of the coaches...in this group have scientific backgrounds. For a stated purpose of transitioning a scientist from academia... -
Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors
TypeBlog PostPublishedAug. 20, 2019, 12:31 p.m....conditions (individually versus in groups), health status, diet, cage enrichment, stress level, origin (vendor...of channel protein, they can trigger neuronal excitation or inhibition upon light (or drug) application...situations such as learning and memory or emotional states, for example anxiety or depression. In upcoming... -
Mouse Modeling, Part 1: Genetically Engineered Mice
TypeBlog PostPublishedJuly 11, 2019, 1:26 p.m....which is then implanted into a female animal for gestation. Nucleases that induce double-stranded DNA breaks...which is then implanted into a female mouse for gestation. All three can introduce multiple base pair deletions...deletions, but CRISPR/Cas9 can also introduce point mutations. While using targeted nucleases to generate an... -
Rabies and Neuronal Tracing
TypeBlog PostPublishedMay 29, 2018, 1:51 p.m....useful for neuronal tracing, it has two practical limitations. First, these high levels of viral replication...an enveloped virus. In general, viral envelopes dictate infection of target cells through their interactions...evolution of inputs onto new neurons in the adult dentate gyrus and olfactory bulb. Proc Natl Acad Sci U ... -
Corralling and Cornering the Goldilocks Antibody
TypeBlog PostPublishedAug. 9, 2022, 1:15 p.m....review analyzing and explaining the antibody crisis states that up to 50% of all commercially available antibodies... technology, it is important to show through qualitative data that they are a long-term solution to reproducibility... above) and literature suggest, providing an expectation and comparison on the antibody signal’s location... -
Antibodies 101: Stripping and Reprobing Western Blots
TypeBlog PostPublishedAug. 1, 2024, 1:15 p.m....respectively. Chromogenic detection produces colored precipitates that permanently stain the membrane, making .... Cover the membrane in stripping solution and agitate for 5–30 minutes at room temperature. The incubation.... Cover the membrane in stripping solution and agitate for 30–45 minutes at 50 °C. Wash the membrane for... -
Progress Towards a PAM-Free CRISPR
TypeBlog PostPublishedMarch 7, 2024, 2:39 p.m.... if you want to knock out a gene, a frameshift mutation introduced anywhere within the early coding region... hand, if you are trying to knock-in a patient mutation or need to make a precise amino acid change, there...away from a desired site does not have the same devastating impact on efficiency as 10-15 bps away does. ... -
High Complexity Golden Gate Assembly with NEB’s DAD
TypeBlog PostPublishedAug. 1, 2023, 1:15 p.m....fidelity. NEBridge GetSet Need a set of overhangs, stat? GetSet can do that for you, complete with predicted... °C cycling 4-base Bsal-HFv2® 37 °C Static 4-base Bsal-HFv2, 1x NEBridge Ligase MM ...They first compared 10-piece linear (the native state of the T7 genome) and circular assemblies. While... -
Design Tips for Prime Editing
TypeBlog PostPublishedJan. 23, 2025, 2:15 p.m....with BioRender.com. Add silent mutations near point mutations to create 3-base (or longer) tracts...’s all our tips for today! Each tip includes a citation to the paper it was drawn from. If we’ve piqued... -
CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9
TypeBlog PostPublishedNov. 14, 2024, 2:15 p.m....the majority of the lncRNA locus, scientists can mutate the transcription start site, splice sites, exons...function (Hazan and Bester 2021; Zibitt et al. 2021). Mutating splice sites is particularly effective as it can... Transcription start site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog PostPublishedApril 6, 2021, 12:30 p.m....Depositing labs often provide insert sequences or annotated Genbank files that are useful for more complicated...genes with many modifications. We look for point mutations, truncations, and insertions that could compromise...compromise function. When we do find mutations, we check to see if they affect the translated amino acid. ... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog PostPublishedAug. 30, 2016, 2:30 p.m....creativity and expertise of the group and yet never hesitated to reach out to the greater research community...portions of the Zika virus genome with Addgene to facilitate rapid sharing of these tools with the Zika virus...and the School of Molecular Sciences at Arizona State University. His group carries out research in synthetic... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog PostPublishedDec. 1, 2015, 3:30 p.m.... the NIH, started off with the first keynote presentation. He talked about the impact of the Human Genome...seeing tRNAs because they contain “hard stop” mutations. New techniques used by the Lowe lab have allowed...species for genome engineering. The final keynote presentation was by Nobel Laureate Thomas Steitz of Yale.... -
7 Simple Strategies to Resolve Conflicts with Difficult Supervisors
TypeBlog PostPublishedJan. 21, 2015, 1:30 p.m....difficult person: 1) Understand your supervisor’s expectations Every supervisor has their own management style...frequently. The easiest way to meet your supervisor’s expectations is to ask upfront: “How frequently would you... details of your work. Describe the problem by stating the facts and explain how it is interfering with... -
Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)
TypeBlog PostPublishedJune 5, 2014, 3:32 p.m....genetically modified organisms, and GMOs have a bad reputation. The public is can be fearful of this technology...ended up applying to a business accelerator freely stating that we had an idea, knew how to build it, but ...in 2004, she accepted a commission in the United States Air Force. She left active duty to pursue a degree... -
Viral Vectors 101: Producing Your rAAV
TypeBlog PostPublishedJuly 16, 2024, 1:15 p.m....like adenovirus (Ad) or herpes simplex virus to facilitate efficient replication. The most common production...immunosorbent assay (ELISA) (Grimm et al., 1999), quantitative PCR (qPCR) (Aurnhammer et al., 2012), droplet...what Addgene uses, check out our ddPCR for AAV quantitation blog post. We generally do not recommend using... -
Twenty Years of Addgene Sharing: CRISPR
TypeBlog PostPublishedApril 18, 2024, 1:15 p.m....our requestors. However, we can again see quick adaptation and steady use of all types of libraries, once...categorized by year of deposit. Outlines show a representative proportion of CRISPR plasmids requested, categorized...total of 23,006 times, including 14,677 unique citations. CRISPR is a story that centers around the impact... -
Transferable Skills Guide: Managing a Team
TypeBlog PostPublishedJan. 30, 2018, 2:42 p.m.... clearly As a manager you need to clearly state expectations and goals. Yours and your team’s success ... schedule a meeting to discuss the work, set expectations, answer any questions, and (most importantly... -
Fluorescent Tagging of Endogenous Genes with SapTrap
TypeBlog PostPublishedOct. 12, 2017, 2:26 p.m.... of many cellular mechanisms, there are some limitations to working with FPs. Overexpression of fluorescently...of normal protein function, and ultimately misinterpretation of the protein’s cellular role. One way to...However, CRISPR protein tagging does have its limitations, especially when trying to perform high-throughput...