...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...

...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...

...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 ...

...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...

...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, ...

... 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...

...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...

...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 ...

...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...

... 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...

...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...

...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...

...participants could share their notes together. Figure 1: Can you draw Spongebob from these instructions without...on Twitter where each poster presenter had four “slides” to use for their poster, eliminating the need ...

...methods to improve AAV production.     References 1.Clark, K. Reed. "Recent advances in recombinant adeno-associated...if you end up concentrating your AAV above your desired titer and want to dilute it, err on the side of...less buffer than you would to achieve the exact desired concentration to avoid over-diluting, and make ...

...loci in the genome and the guides are relatively unstable.   Figure 1: CRISPRainbow labeling. Two... the gRNA tetraloop and went through subsequent design iterations to increase stability and minimize misfolding...

...regarding what makes this group of researchers special: 1. Stem cell scientists care deeply about ethics Ethics...speakers from top minds in the stem cell field have described the Yamanaka paper as "life changing". Dr. Yamanaka...affordable. 4. Stem cell scientists are generous Besides some of the scientists I've already mentioned, ...

... genes with a single viral vector.   Fig. 1: Schematic of multiplex gene editing with DAP array...The natural CRISPR locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to ...applications, we provide detailed manual and online design examples of using Addgene plasmids hCtRNA_FT (#186715...

...as an endogenous RNA imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head ...effects, both of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% ...crystalizing the hfYFP structure, the group rationally designed GFP variants with similar capabilities. If you...

...settings with experimental samples. Figure 1: Example flow plots for a single color control. A)...familiar with the basics of controls in experimental design, you’ll need a few controls specific to flow as...controls like FMO or isotype controls, which we’ll describe a bit later. Single Colors Single color controls...

...Networking 101-Building Relationships for Success”. Choose 1-2 tactics to try. One of my favorites is making the...mentoring questions I set out to answer. First, I will describe some of the many ways you can approach finding...Watch for my upcoming mentoring post in which I describe how to organize a successful peer mentoring circle...