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TypeBlog Post...them you can do today! 10 ways to share your science 1) If you don’t already make it a habit to talk – ...research, SciFund is really motivated more by the desire to help close the widening gap between science ...
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Viral Vectors 101: Transductions
TypeBlog Post...depending on the virus, cell type, and/or desired event. Figure 1: Transduction to expression process...retrovirus, lentivirus, adenovirus, or AAV. Then, your desired DNA to be integrated can be cloned into the vector...packaged, when the vector is expressed. Once the design and construction steps are complete, it’s time ... -
Plasmids 101: Gram Negative and Positive Bacteria
TypeBlog Post...heat-fixed smear with the crystal violet stain for 1 minute. Rinse the slide briefly with tap water. Flood...phrase gram negative or positive being used to describe a species of bacteria, but what does it actually...bacteria. In a gram negative bacteria, the alcohol destroys the outer lipid membrane, and the thin inner peptidoglycan... is also useful in a laboratory setting. When designing a plasmid to be inserted into a specific bacterium... -
Behind-the-scenes of the Isolation of the Thermostable IgnaviCas9 From a Yellowstone Hot Spring
TypeBlog Post...could be kept frozen after collection. Figure 1: Paul included a voltmeter on top of the battery box...and workflows where CRISPR nuclease activity is desired and time or protocol steps can be saved by flexibly... -
What Do I Do Now? Academic v. Non-Academic Career Decisions
TypeBlog Post...occur, but good performers move around easily Table 1: Taking it a Step Further - Big Company v. Small Company...one list can ever encompass them all. We can’t designate non-academic jobs as “alternative” anymore. ...Develop expertise areas quickly Jack/Jane of all trades More sources of training and resources within... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog Post... difficult to modify in the past. Figure 1: Identification and enrichment use of XMAS-TREE. a...without end-point sequencing assays. This tool was designed by putting together the coding sequence of a mCherry...MYC-CRISPR library targets E-boxes genome-wide. It was designed based on MYC-ChIP-seq data from several MYC-dependent... -
Another Pathway into Cells: iTOP
TypeBlog Post...vesicles. Okada CY, Rechsteiner M. Cell. 1982 May;29(1):33-41. PubMed. Protein transduction: unrestricted...transduced using only protein. Read on for a description of the lab’s iTOP protein-only transduction method...transduction by osmocytosis and propanebetaine) to describe this method of protein transduction. Later studies... -
Visualizing Translation at the Single Molecule Level
TypeBlog Post... cell types and physiological states. References 1. Ingolia, Nicholas T., et al. “Genome-wide analysis... determine if proteins are targeted to their destination co- or post-translationally. TRICK: Visualizing... usually translated in a specific compartment. Despite these limitations, these techniques represent powerful... -
Components of CRISPR/Cas9
TypeBlog Post...components necessary to: 1. Bind to a guide RNA The guide RNA, which will be described in more detail below...express a gRNA designed to direct Cas9 to their target sequence of choice (see tips for designing a gRNA) and... of the major CRISPR lingo and will be able to describe the functions of the various CRISPR/Cas9 components...responsible for processing foreign DNA as well as the RNA guides required for endonuclease function, when used for...DNA damage is a DSB within the target DNA ~3-4 nucleotides upstream of the PAM sequence. Cas9 Species/Variants...with the appropriate PAM sequence) to modify the desired genomic locus. Some historical notes CRISPR ... combined with its adjacent spacer, ultimately encodes a single crRNA. The direct repeat regions contain... -
How Addgene Creates Content
TypeBlog Post...to do that than in the Addgene blog? Fig. 1: One of our favorite content collections is our Help...up walls of text. Since our guides are heavily used, our graphic designer made most of the Antibody Guide... quickly." Final design (right). Immediate and immense regret of the graphic designer not shown. ...blog, website, social media, videos, newsletters, guides, and e-books – comes into play. Our goal is to ...communication between our content team and our graphic designer to make sure things are clear, accurate, and in...Reference image (left) provided to our graphic designer upon request that the science communicator "just... -
Behind the scenes: Addgene’s new search engine and more
TypeBlog Post...present in the data such as synapsin, hsyn synapsin-1, human synapsin, hSynap. Matching We capture multiple...educational resources — including blog posts, protocols, guides, and collections — is a lot of material to sift... launched earlier in the year, as well as our redesigned homepage and our new search tool, are the results...a few of the strategies we used in our software design. In each case, we leveraged the team’s knowledge...per resource category to expose as filters and designed the search engine to return only the filters that... bits of information for a given resource, and designed the search engine to weight search results differently...'s new search Read to learn more about why we redesigned our homepage and mascot ... -
Plasmids 101: Methylation and Restriction Enzymes
TypeBlog Post...into account, the digest pattern depicted in Lane 1 would be expected for a XbaI/NotI digest. The program... NEB's Resource describing Dam and Dcm Methylases Discovery of DpnI Review describing dam-/dcm- E. coli...mutagenesis. During this process, incorporation of a desired mutation into your plasmid of interest by PCR generates...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which... -
Scientists Map the SARS-CoV-2-Human Interaction Network
TypeBlog Post...faster. Years of science, done in weeks Figure 1: Sending out 80 FedEx envelopes in a single day --...s time, we’ve published an article on BioRxiv describing a comprehensive SARS-CoV-2 virus-human interaction... viral proteins except for Nsp3 and Nsp16, and includes an additional catalytically dead version of the...the viral protease, Nsp5. Our set also includes a GFP construct as a control. For easier portability, ... -
Communicating Your Science With Help From ComSciCon
TypeBlog Post... will be a positive asset throughout your career? 1. Make scicomm practice a priority The good news for... craft sentences for abstracts and presentation slides, learn from audiences as they do science outreach...about whether the same principle applies to other slides in your deck. If someone asks a question about...of a new science communication initiative that provides yet more scientists with opportunities to practice... -
Plasmids 101: Blue-white Screening
TypeBlog Post...and also by a lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate...bacterial lac operon contains a gene called lacZ that encodes for the enzyme β-galactosidase. Expression of the...process is called α-complementation. The system described above was put to practical use in the following...Although blue-white screening is probably the most widespread way to select for plasmids containing an insert... -
Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools
TypeBlog Post...extension. Now, they split EB1 into two halves (Fig. 1): one containing the microtubule binding domain and...colleagues started with the FP mNeptune684 (originally described by Li ZY, et al. PLoS One, 2016) and tested a ... -
Plasmids 101: Origin of Replication
TypeBlog Post...produce as many as 700 copies per cell. Figure 1: A plasmid map showing the standard features of a ... A -C compatibility grouping is an arbitrary designation, and plasmids from the same incompatibility group... -
Advice for Moving Into Sales After Your Science Postdoc
TypeBlog Post... After all I was only going to be interviewed for 1 hour so having a broad enough insight was going to...Coordinator’ came up at Abcam. Unlike other job descriptions for marketing positions posted on recruitment... -
Hot Plasmids: FRET-Based Biosensors
TypeBlog Post...variants. This resulted in the construction of redCALWY-1, a red-shifted FRET sensor for zinc using variants...beginnings of the field. Those engineered sensors were designed to replace synthetic calcium dyes, which had been... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog Post...can also rank the plasmids by abundance, scaled to 1, to create a “cumulative wealth distribution” curve...next-generation sequencing (NGS). For gRNA libraries, you’ll design primers to create 200-300 bp sequencing products...