...Almo, Institute for Protein Innovation  1:18:45 - 2:01:18 (end) - Panel Alex Tucker (Ginkgo Bioworks)...their curation of citable, linkable figures. Cause #2: Hidden “negative” data We all want to be able to ...experimental designs and we need to be transparent about the reasons we alter these designs if we do so...experimental design to a journal prior to performing the experiment (or set of experiments), the design would...from the event can be found below along with a description of the program beneath it. You can jump to the...the different time stamps in the description to watch any sections you’re particularly interested in, ...JoVe, and Bio-protocol all work to provide more descriptive and interactive protocols that make it much easier...

...fusion location, there are many choices to make. 2. Prime Editing: Adding Precision and Flexibility to...on the Addgene blog and have created and updated guides on various topics in biology over the past year...containing these fluorescent proteins at Addgene. 7. Designing Your Chalk Talk for the Academic Job Interview...

... editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...cytosine and adenine base editing, called CABE-Ts and TadDEs without additional fused components (Lam et al....

...PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary T. Britton, and Anne S...subtilis and Pichia pastoris pCri also includes plasmids designed for protein expression in systems other...construct, most often in special E. coli strains designed for protein expression. Cultures are then lysed...plasmids are divided into two sets, each of which includes plasmids with different varieties of tags, both...other eukaryotes better than E. coli. Plasmids designed for these systems may include a signal peptide...can improve disulfide bond formation. pCri also includes a special feature for difficult-to-purify membrane...membrane proteins. pCri vector pCri-13a, designed for use in B. subtilis, contains the MISTIC (membrane-...

...having trouble preparing your most excellent resume. 2) Follow job boards and check them regularly Use your...experience. Most Addgene PhD level job descriptions clearly describe that the roles do not involve bench ...submission of a job application. 3) Read job descriptions carefully Be open minded and be bold. You don...qualifications to apply for a position.  A job description is usually a wish list and the hiring manager... way to a hiring manager. Learn to confidently describe your strengths in your resume and don't be shy...

...system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper... a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using...protein analyses using ELISA. Peptides 72:4–15 . https://doi.org/10.1016/j.peptides.2015.04.012 Burnette WN ...blocking buffers, and incubation times. If the vendor includes publications that cited the antibody, consider...abundance In addition, the thickness of the band provides information about the relative abundance of the...

...teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...application process so you can learn more! Figure 2: The many ways to prepare for a science policy career...uncertainty of it all. Though I had spent the first few decades of my life following a very straightforward conveyor... harassment in academia or a large company who decides how they are going pursue regulatory approval for...their Local Engagement Playbook for step-by-step guides on how to engage in your local community.  Read...comprehensive resource developed by Dr. Kelly Singel which provides an overview of all available science policy fellowships...

...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...to be present and recognizable by your antibody despite the manipulation. Additionally, not all sample ...large deletion that removes the entire exon that codes for the epitope recognized by your antibody. You...specific, then you expect that the bulk of the peptides identified to be from the target protein. This...tuned for ready-to-use antibodies from Addgene Designing a recombinant validation experiment? Find plasmids...

...melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo, Rochelle...wall of green algae (Soo et al., 2015). There, it divides into two cells. At the same time, it sucks out ...1983). Grasshopper parasite The parasitic worm Spinochordodes tellinii‘s larvae makes its home inside grasshoppers...

...former lab at UNC Chapel Hill. Bähler et al.(2) describe a modular collection of plasmids for a wide variety...protein products is desired, as multiple selection markers can ensure that all desired tags have been integrated...integrated. Simply design your amplification primers with the desired targeting homology—in frame, of ...These tagging vectors are based on previously described pFA6a-link vectors and include a Kan, SpHIS5, ...3xHA, 13xMyc, GST, or GFP). Longtine et al.(3) describe a complimentary set of plasmids for use in S. ...

... and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed Repair...problem - see our recap below! Day 1: gRNA design tools: How to Design your gRNA for CRISPR Genome Editing and...given our CRISPR Plasmids and Resources a fresh new design, sorting the plasmids we have available by category...capabilities. Epigenetics: CRISPR has made it possible to design programmable epigenome editing tools, and the field...cleavage in bacteria. In mammalian systems, Cas13a degrades only the specified RNA target. Bonus: It displays...

...might otherwise be hard to identify. Figure 2: Users can cross-interrogate screens from the literature...ve just discovered that your favorite gene was described in a CRISPR screen publication. You see this mentioned...This format is inherently flexible and allows the broadest possible scope of screens to be curated, for example...other than cell viability. The following video provides an in-depth walk through of how to use ORCS:   ...

...1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to maximize activity and ...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini... massive amount of descriptive transcriptomics data. Custom CRISPR screens designed to target differentially...Institutet, in Sweden. It can be very time consuming to design 5 guide RNAs (gRNAs) targeting each of the 1000...). Green Listed is a new software tool used to design gRNAs for custom CRISPR screens targeting a (long...moment, 13 full genome SpCas9 knockout libraries designed by different academic labs, targeting mouse, human...drug targets  Another interesting approach is to design screens against known druggable targets, preferentially...

...09656-2 More resources on the Addgene blog CRISPR101: Validating Your Genome Edit How to Design Your gRNA...LaFleur, et al. 2024.   The X-CHIME deposit also includes pXPR_071, a plasmid that leads to higher titer...

...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available pCAMBIA...Improving Agrobacterium binary vectors Vectors designed for use in Agrobacterium are termed binary vectors...ideal for large gene transfer. The M24 promoter provides constitutive, high-level gene expression and is...

...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ... favors the breakage of the hydrogen bonds, destabilizing the molecule and favoring the separation of ...nuclease free water is a great option because it provides a great versatility on how to use your plasmid...

...E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference between arg1 and arg2 vesicles...have hollow interiors and a protein shell that excludes water but is gas permeable. These vesicles are...sequentially exposed to acoustic pulses with high enough amplitudes to first collapse arg2 and then arg1 vesicles...

...closer to the clinic (Tsai and Joung 2016). Table 2: Unbiased Genotyping Options Technique Detection...PMID: 23792628. PubMed Central PMCID: PMC3773023. 2. Vouillot, Léna, Aurore Thélie, and Nicolas Pollet...good starting point for most researchers as it provides a list of putative off-target sites that they ... approach misses larger mismatches (e.g. six nucleotides) that may still lead to off-target double-stranded...contains both the rare off target event and the desired edit is low. Therefore, unbiased sequencing may...

...access this tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool.   DNA sequence...primer design Design primers for Goldengate cloning   Design primers for In-Fusion Cloning Design primers...traces, while the paid version provides more tools for plasmid mapping and design (Figure 1). Benchling: While...academic users. Serial Cloner: Free desktop-based software for plasmid design and mapping. ApE (A plasmid Editor...Primer design tools Many of the above-listed plasmid mapping tools have built-in primer design tools, ...stand-alone sites for general primer design, as well as primer design for different cloning applications...characteristics. NEB Data-optimized Assembly Design (DAD): This tool designs overhang sets for high complexity Golden...

...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout... whose expression varies depending on transgene design and delivery. They generated hPSC lines with AAVS1...