Virus Protocol - Generating Stable Cell Lines
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Protocol
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Improving Transduction Efficiency with Exosome AAVs
TypeBlog Post...the widely used 293T. During vector production, assembled AAV vector particles accumulate inside the producer... are often found with cellular exosomes, small membrane vesicles or microvesicles (approx. 40 - 150 nm... diameter) that are secreted from the cellular membrane into the culture medium. AAV particles are either...found that the association of AAV particles with membrane vesicles protects them from antibody recognition... -
Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**
TypeBlog Post...by incubating embryos in a MG-2I solution, or injecting adult fish with the dye. Embryos exposed to just...experiments with MG-2I-dL5** were performed with embryos and adult fish. Zebrafish are an ideal model organism...organism for these experiments because developing embryos are transparent, so it’s easy to watch development... -
Addgene Welcomes John Doench to the Advisory Board
TypeBlog Post...The Addgene Advisory Board consists of prominent members of the bioscience and tech transfer communities...apply to graduate school at MIT. Doench excitedly remembers: “When I came to MIT to interview, part of the...process was several short talks by current faculty members and it just blew me away…. I have to come here...Find CRISPR pooled libraries Learn about other members of the Addgene Advisory Board Additional Resources... -
Management for Scientists: Delegating is Key
TypeBlog Post...ideas, initiative and creative thinking. The team members will be more invested in the projects and outcomes... tasks are distributed appropriately. If team members have opportunities to develop and stretch then ...then facilitate appropriate communication between members of the team to ensure that no component of the ...management style. Recipes for delegation success Remember that there are always things that the manager ... -
Finding and Joining Your Dream Lab
TypeBlog Post... hires initially based on those who notice and embrace the “odd and extreme openness” of the lab (i.e ...interests, and spends one-on-one time with the current members to share short-term goals and chat about awesome...questions for a candidate to ask lab heads, lab members and lab neighbors during the interview process....alumni taken diverse career paths? Ask other lab members how the advisor treats such inquiries. If you know... -
Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification
TypeBlog Post...let the team detect its co-localization with a membrane-localized mCherry or GFP protein. With this system...bind, it co-localizes with its target at the cell membrane and when the OptoNB can’t bind, it localizes to...proliferation and differentiation. Since Ras is a membrane localized GTPase, the team fused a mCherry OptoNB...OptoNB-SOS fusion protein in cells that expressed membrane-localized mCherry. With darkness, SOS binds Ras... -
Viral Vectors 101: Viral Vector Elements
TypeBlog Post...pseudotyped to alter infectivity. Next, the virus must assemble and have appropriate factors displayed on the ...nucleocapsid viral components needed for viral packaging, assembly, and display. Pol, the last component, contains...structural capsid protein that helps with virion assembly and particle release from late endosomes when ...viruses discussed. LXs/Iva2 are primarily capsid assembly proteins required for packaging of the virus into... -
Sharing is Caring: But How to Distribute Open Hardware?
TypeBlog Post...material (BOM) of an assembly, the feasibility of manufacturing, and subsequent assembly can be difficult ... a similar way to the biological protocol, and assembly designs based around the use of commercially available...developed techniques are the key enablers for making assemblies open-source. The sharing of digital representations... -
Developing a cold tolerant E. coli using specialized chaperones
TypeBlog Post...were designed by our team member Kallol Mozumdar. Sreepadmanabh M is a member of the team from IISER Bhopal...This post was contributed by Sreepadmanabh M, a member of the iGEM team from IISER Bhopal. Following up...in extensive outreach programs to encourage and embolden the spirit of scientific innovation amongst school... -
Viral Vector Guides and Plasmids
TypeCollection...the necessary components of the viral genome to assemble the viral capsids (packaging and envelope plasmids... -
Supporting Rare Disease Research with the 2020 BeHEARD Awards
TypeBlog Post...interested in TS when I was a newly independent faculty member studying sex differences in thoracic aortic disease...emphasis on oligomerization, signalling complex assembly, and localization to the nucleus,” says Dr. Terry...consist of many parts that work together on the cell membrane. Germline mutations in one of these parts, the...to Addgene. Congratulations Terry Hébert! Members of Terry Hébert's lab. Additional Resources ... -
Transferable Skills Guide: Public Speaking
TypeBlog Post...quickly string together all the words that I could remember related to this paper and/or biology in general...perspective. Most people won't notice, or won't remember and those who do are very likely to be sympathetic... diving board charades, I doubt anyone but me remembers any of the incidents above–and the dumpster noises...if it doesn’t go perfectly,” can help. (And do remember to breathe!) Public speaking as a transferable... -
Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics
TypeBlog Post...-opsin GEVIs. With an increase in the neuron’s membrane potential, Ace absorbance increases and results...biologists to use Type IIS restriction sites to easily assemble multiple DNA fragments. This system has been used... for greater reproducibility as scientists can assemble parts from different labs and experimental systems...biology tools for cyanobacteria. Figure 3: Assembly of plasmids using the Cyanogate Kit and subsequent... -
Uncertainty about Labor Law Brings More Uncertainty to Postdoc Wages
TypeBlog Post..., executive director of Future of Research. On December 1st 2016, many postdocs working more than 40 hours...recently in a paper published in F1000Research on November 2016. Due to the nature of the postdoc position...been made publicly available (below). However, on November 22nd a preliminary injunction against the FLSA...for their postdoctoral salary information as of December 1st. Salary inconsistencies across geographical... -
Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells
TypeBlog Post...multimerize and spontaneously assemble into particles at the cell membrane. Mature Nanoblades then bud ...CRISPR to multiple types of targets: primary cells, embryos, and animals. Nanoblades can also carry different... where they can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote... -
Mouse Modeling, Part 2: Breeding and Crossing Mice
TypeBlog Post... to introduce genetic modifications into mouse embryos. But once you generate a growing colony of genetically... as more mutations are introduced into a single embryo, the likelihood that a mouse will end up with the... are homozygous for all of your desired genes. Remember Punnett squares from high school biology? They...transgenic mice that can impact reproduction include embryonic lethality, a short lifespan, or infertility and... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post...If you were born before 1985, you might remember going to the store and buying CDs when you wanted to...antibody and when translated into protein will assemble into a fully functional antibody. These antibodies...antibody are encoded in a plasmid. (Right) An assembled antibody protein binds an antigen. Image from ...L, Xin L, Shan B, Li M (2016) Complete De Novo Assembly of Monoclonal Antibody Sequences. Sci Rep 6: .... -
Pouring LB Agar Plates
TypeProtocol...with your antibiotic of interest. Last Upload: September 14, 2016 Protocol Video Before you begin this ... -
Plasmid Cloning by PCR (with Protocols)
TypeProtocol...to ligate our cDNA into the recipient plasmid. Remember to insert your DNA in the correct orientation ...