... both substantial merit and national importance; (2) The foreign national must be well positioned to advance...outstanding achievements in your specific academic field, (2) that you have three 3 years of research experience..., Casseus Law suggests a mix of experts including 2 dependents –people who have worked with you i.e. your...The recently established case Matter of Dhanasar provides the three-prong test that US Immigration officers.... As Immigration is a federal law, Casseus Law provides legal services to people located in all 50 states...

...tagging." Nature methods (2015). PubMed PMID:26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and...First described in the 1980s, protein tags are now one of the most useful items in a scientist’s toolbox...IT10 expressed well in cells and did not form undesirable aggregates. Using budding yeast as a model system...

...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...replicate—but not always to the same extent. As described in our previous Origin of Replication post, DNA...replication. For example, the plasmid ColIb-P9 encodes the protein RepZ to drive its own replication. ...

...efficiencies 2-3-fold. However, the process of double nicking increases indel formation slightly. Designing the... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...site by the pegRNA — a guide RNA that also encodes the desired edit and homology to the genomic DNA locus...both specifies the target site and contains the desired edit (or edits) engages the prime editor protein...and the reverse transcriptase domain copies the desired edit, directly polymerizing new DNA onto the nicked...edit within what Fyodor Urnov refers to as “PAM deserts” in the genome. More versatile and precise than...

...Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One Step, Precision...colleagues describe in the PLoS ONE paper that introduced the kit in December, GreenGate includes the six... bring this into your target organism. We have designed a Golden Gate system based on the daily needs ...and cut another,” Lohmann said. “It allows us to design modules that always have the same recognition site...produced will have different overhangs, they can be designed such that they go back together only one way and...

...publication at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene...Specifically this includes primer directed isothermal RNA amplification, new toehold switch designs, CRISPR/Cas9...is an Assistant professor at the Biodesign Center for Molecular Design and Biomimetics and the School of...development of diagnostics. Mostly transmitted by Aedes mosquitoes (aegypti and albopictus) and, aided by... toehold switches, which allow for the rational design of RNA sensors that can detect virtually any RNA...areas conducive to the spread of the virus by the Aedes mosquito (Messina et al, 2016). We quickly assembled...spread out the risk for the project by initially designing and ordering several dozen versions of the toehold...

...at Addgene – 500,000 plasmids shipped! That's a 1/2 million plasmids sent to happy scientists around the...information in our lab software, labeling stabs with barcodes as they go. Stabs are organized by shipment and...

...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...light-inducible transcriptional effectors (LITEs) designed to bind specific genes and turn them on or off...News & Views Andreas Möglich and Peter Hegemann described several advantages of the new system: "First,...

...  Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic of Opto-RhoGEF design and activity...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...optogenetics than neuronal activity. Researchers have designed a variety of photoswitchable elements into many... and limitations – for example, microtubule-destabilizing drugs are impossible to target to specific subcellular...plasmids if you want to make a stable cell line. This design allows highly localized, rapid and reversible microtubule...processes in cells and tissues. And, this Opto-RhoGEF design could be extended to other GTPases. The Chow Lab...plasma membrane. In previous publications, they described tools to control formation of lamellipodia and...

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...comments section.  FX cloning kit This kit is designed to be used with Type IIS restriction enzymes and...this kit have been used to purify a variety of peptides including antibiotic efflux pumps and ankyrin ...et al. "A transcription blocker isolated from a designed repeat protein combinatorial library by in vivo...followed by leader sequences (encoding small leader peptides) downstream of which you can clone your gene of...Shine-Delgarno sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to...

...would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for...gating on the negative population.     Figure 2: Dump gating labels all unwanted cell lineages with...when it comes to fluorophore selection for panel design. Find streptavidin plasmids and biotin plasmids... this system is increased flexibility for panel design. Another perk is that one primary antibody can ...for example, a previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside analogs...walk through the topics we covered in this post. Besides direct labeling, indirect labeling can be used ...surface, dump gating is a useful way to get rid of undesired cell populations. Dump gating uses the same color...

... used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be ... 296.5569 (2002): 913-916. PubMed PMID: 11988576. 2. Tsien, Roger Y. "The green fluorescent protein." ...cofactor binding proteins as described above. The smallest tags are peptides that are engineered to bind...Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains...

...Public Release Event scheduled for March 4th from 2:30-4:00 pm ET.  You can learn more about the report's...job of providing skills training for scientists destined to continue in academia. It seems it’s largely...

... on the future of the scientific endeavor October 2-3, 2014 Boston University Jacob Sleeper Auditorium...work with any group of trainees interested in designing career exploration programs or peer mentoring ...

...SnapGene Viewer (Mac/PC) FinchTV(Mac/PC) Chromas (PC) 2. You should see individual, sharp and evenly spaced...are usually unresolved and small, so I suggest designing your primer at least 50bp upstream of the sequence...

...similar optical quality (magnification of 140X and 2 micron resolution). I have worked many hours in a ...is absolutely fascinating to see the living art designed by creative minds with a (I assume) very steady...Our Microbiology Resource Page CRISPR plasmids designed for use in bacteria Deposit Your Microbiology ...

...favorite molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique to screen...record breaking month and to find other posts that deserve a second look. 5 Great Apps for Lab Life This very...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ...Jessica Sacher from the University of Alberta and describes a technique you can use to find phage proteins...that, while affecting a small number of people, deserves our attention and has huge potential for the development...

...that you can easily spot the colors and text. Tip 2: Use electronic to-do lists to access your tasks on...lab earlier, to read that stack of papers on your desk, and to tackle some new experiments before the next...gathering that pile of lab notebooks and binder on your desk. Arrange them in chronological order (or by project...labels. Color code your Post-its and write brief descriptions on each. Place the notes sticking up out of ...colleagues. Start small by identifying the protocols, guides, or references that your labmates ask to borrow...

...develop therapies to treat disease in the lung. Chapter 2: Balancing research with medical practice as an MD...University Center for Regenerative Medicine. Darrell describes his career goals and what drives him as a praticing...

...access to the results of federally-funded research. 2)      In June, the White House honored 13 “Champions...     Science’s John Bohannon published an article describing a sting operation in which he submitted a phony...

...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. ...