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TypeBlog PostPublishedOct. 11, 2018, 12:30 p.m.... DNA fragments in a single “one-pot” reaction to form complex, multi-insert modular assemblies that enable...ligase. Inserts, either precloned or in amplicon form (shown above) are flanked by BsaI sites in an orientation...number of transformants); the greater the number of inserts, the lower the number of transformants. To compensate...containing the transformants onto the selection plate. Figure 2 shows representative transformation plates obtained...throughput cloning efficiency measurements. We performed additional efficiency measurements using similarly...background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion of the...of the 5-, 12- or 24-fragment test systems was performed by setting up assembly reactions in which a single...
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Plasmids 101: Secondary Nanobody Toolbox
TypeBlog PostPublishedFeb. 27, 2018, 2:04 p.m....antibodies cannot be incubated together without forming large oligomeric complexes. Antibodies are also...Maleimide-labeling occurs when a maleimide group forms a covalent bond with a sulfhydryl group (–SH), such...conjugation to a specific label (see step 4 for more information). Purify the Nanobody With A Nickel Column: Lyse... -
Deciphering the Mysteries of Behavior with Viral Vectors
TypeBlog PostPublishedSept. 17, 2019, 12:40 p.m....norepinephrine release causes a freezing response, a form of immobility caused by stress and fear. Such a ... be used as behavioral readout to study certain forms of drug addiction. When an animal has been exposed...discussed some of the general considerations for performing behavioral tests, but now we’ll focus on how ... -
How to Write a Scientific Review Article
TypeBlog PostPublishedFeb. 16, 2017, 3:30 p.m....down on the page as possible. This could be in the form of lists, streams of consciousness, or anything ...most useful parts of reviews. These could be in the form of figures/schematics or tables. And don’t forget...literature, and to synthesize a large volume of information: basically, a scientist’s tool kit. I wrote a...will be a temptation to include every piece of information that was ever published. Obviously this isn’t...accord, once you have these rules (word limit, formatting guidelines, etc.) you have some criteria to shape...the journal requires. Using the (Last name, Year) format also has the benefit of exposing you to relevant...researchers in the field. I made sure to find information from genome-wide studies, as well as results ... -
Our New Antibody Mascot is....
TypeBlog PostPublishedFeb. 24, 2022, 2:15 p.m....help us decide by voting for your favorite on this form or head over to Twitter to vote on @addgene's Twitter... -
Addgene’s Newest Way to Speed Your Research: The Cloning Grade DNA Service
TypeBlog PostPublishedJuly 28, 2020, 1:15 p.m....the future using the Cloning Grade DNA Suggestion Form. Additional resources on the Addgene blog Browse...you could directly start your digest, PCR, or transformation? Great news. A subset of our collection has... -
New Podcast Segment: Hot Plasmids
TypeBlog PostPublishedMay 17, 2017, 3:37 p.m....Our Segment on CRISPR X Hot Plasmid 2: Magneto 2.0 form Ali Guler's Lab - Control neuronal activity with... -
Editor's Choice, July 2016
TypeBlog PostPublishedAug. 5, 2016, 3 p.m....interested in blogging for us, fill out our Guest Blogger Form and we'll be in contact with you ASAP. Happy Reading... -
Using AAV for Neuronal Tracing
TypeBlog PostPublishedAug. 9, 2018, 1:04 p.m....reciprocal, forming a neuronal loop which is activated and strengthened during memory formation and memory...neuronal cells. Synapses at the terminals of axons form connections to dendrites of proximal neuronal cells...on kinesin. Different rates of speed for various forms of retrograde and anterograde transport suggest ...trafficking are not fully understood and may be a form of transcytosis where the AAV particle is taken ...dendrites receiving incoming information, and a projecting axon sending information onwards to other neuronal...prefrontal cortex brain regions is involved in the formation of episodic memory, a special type of memory which...(see Jin & Maren, 2015). Directional flow of information between different parts of the brain is mediated... -
22 Hot Plasmid Technologies from 2014
TypeBlog PostPublishedJan. 6, 2015, 5:21 p.m.... shift occurs during Timer protein life due to a form of oxidation (dehydrogenation). MitoTimers are composed...Protein phosphorylation is one of the most abundant forms of post-translational modifications in cells and...system, the Rinehart lab synthesized the activated form of human mitogen-activated ERK activating kinase...similarly assembled into a Level 2 vector, thus forming a functional genetic circuit. Flexibility has been...amplication reactions for recombination in SLIC cloning to form the desired vector. The pCoofy vectors contain the..., watch informational videos, download protocols, and more! Looking for background information on CRISPR... your experiments. For more information, please see Addgene’s information page for the Rinehart phosphoprotein... -
Newly Updated AAV Data Hub!
TypeBlog PostPublishedFeb. 7, 2023, 2:15 p.m....Hub! The process is easy with our new submission form - and no worries if you get interrupted by a freezer...spread and cellular tropism* (min. 300 dpi, PNG format) At least two replicates of your experiment* Functional... -
Congratulations, Deck The Lab winners!
TypeBlog PostPublishedJan. 14, 2021, 1:30 p.m....since snowflakes and viruses both exhibit similar forms of symmetry. 🎶Walking in a Winter Wonderlab🎶 ... -
Editor's Choice, October 2016
TypeBlog PostPublishedNov. 4, 2016, 2:30 p.m....school much easier to manage. This episode takes the form of a discussion between myself and two of my previous... post this month shares incredibly important information for anyone considering a postdoc. In “Changing... -
Twenty Years of Sharing: Expanding Accessibility
TypeBlog PostPublishedNov. 21, 2024, 10:47 p.m....issue was found in a less tangible form: knowledge and information about the materials in our repository...that our shipping team is always engaged in some form of problem-solving. When universal solutions aren...help with sharing information through the use of popular Chinese social media platforms. As you can imagine...Protocols Agarose Gel Electrophoresis Bacterial Transformation How to Design a Primer Creating Bacterial Glycerol...series even more accessible by collecting and formatting all 101s on a particular topic into beautifully...that lets us know that Addgene’s materials and information are truly accessible, not just available. Looking...repository, from physical materials to useful information, but we’re not done yet… in fact, we may never... -
CRISPR 101: Homology Directed Repair
TypeBlog PostPublishedJan. 26, 2023, 2:15 p.m....are four-stranded branched structures that form when elongation of the invasive strand “captures” and synthesizes...synthesis to fill in the resulting ssDNA. A single HJ forms during this process and resolves by cleavage of ...repair off the other strand; this results in the formation of a DNA structure referred to as the displacement...DNA repair synthesis, ultimately leading to the formation of double Holliday junctions (dHJs). dHJs are ...in DSBR, following strand invasion and D loop formation, the newly synthesized portion of the invasive...additional resources below for more in-depth information on HDR and CRISPR. Our CRISPR 101 series was ... -
Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names
TypeBlog PostPublishedApril 2, 2019, 1:44 p.m....hedgehog, the denticles actually clump together forming one patch on the surface, giving the embryo a “...visceral mesoderm and subsequently the gut does not form properly (Azplazu and Frasch, 1993). I could not...SHH plays a key role in proper limb and digit formation as well as neural development. Specifically, SHH...1993). Tinman is specifically involved in the formation of the heart amongst other things. Rolf Bodmer...Bagpipe on the other hand, is important in the formation of the visceral mesoderm which differentiates ... -
Replacing Paper: Tips for Choosing an Electronic lab Notebook
TypeBlog PostPublishedJuly 10, 2018, 1:08 p.m....as in industry. The news is good. In their ideal form, ELNs provide users with a holistic means to manage...dates of your actions. These records can take the form of written comments, added files, notes etc. Detailed...currently exchanging information with your team? If you need to exchange information regarding lab work ...paper notebooks. Instead, all of the important information is consolidated to one place, easily retrievable...ELN should allow you to collaborate within the platform and avoid countless emails. Important collaborative...permission that only enable them to see the necessary information. Does your lab need to meet any specific compliance...Detailed records about every action performed in the lab (user, date, time, action) are required for regulatory... -
What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program
TypeBlog PostPublishedMay 2, 2019, 12:41 p.m.... time. I was already a biology geek, but this formative experience nailed down my plan to become a research...There is diversity in the social bonds that they form including genetic mechanisms that control who a ...species. Discovering ways to culture, propagate, transform, store, and share all of these new organisms continues...engage in ongoing, real-time dissemination of information and materials. Many groups publicized workshops... or Prochlorococcus with your next project. Gymnotiformes and Mormyridae (Jason Gallant) Weakly electric...mushrooms) but there are over 1,000 species. No transformation techniques are available yet. Polistes (Amy... a system using novel vesicles to efficiently transform these cells which don’t like foreign DNA at all... -
10 Steps to a Perfect Science Talk
TypeBlog PostPublishedAug. 23, 2016, 2:30 p.m....remember better when we hear material delivered in the form of a story. Using stories is one of the best ways... suggest that all graduate students and postdocs form peer mentoring groups to practice giving talks and...of experimental results in the order they were performed does not make for a memorable presentation anywhere...same topics many times, but I always seek out information about the particular audience and venue before...reading the explanation. The people that need the information will find it. Experienced speakers present useful... a crowd. Having answer slides ready after your formal conclusion slide will also help with your confidence...with other ways to engage the audience. Take an informal poll by asking for a show of hands on a particular... -
Site Directed Mutagenesis by PCR
TypeBlog PostPublishedAug. 2, 2016, 2:30 p.m....extension can usually ensure that the 3’-base(s) do not form secondary structures. The introduction (or ablation...plasmid (usually from incomplete DpnI digest) can also form colonies under these conditions. Screening If a ...constructs), in functional analysis of proteins (to perform alanine scanning mutagenesis or targeted substitution...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked plasmid (the...the adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required...required since the E. coli you transform your PCR products into will efficiently patch up the DNA. The ...parental plasmid provides a mean for selecting for transformants which have taken up the mutagenized plasmid....