...: How did you end up becoming a graduate student? 4:50 - 8:00 : How did you approach doing rotations? ...graduate school community to help keep you excited about coming into lab even when things get tough in the...

...description of the experimental conditions.     Figure 4: A screenshot of the materials and methods section... antibody worked under the conditions described above, and a short description of how the antibody performed...

... viruses in a BSL-2 lab instead of a BSL-3 or BSL-4 lab. For more details, check out this review article... with a deletion-mutant rabies virus. Nat Methods 4:47–49 . https://doi.org/10.1038/nmeth999 Wickersham... also toxic at high concentrations as described above and is inactivated by the complement system in serum...

...preferably near the 5’-end) and that also generates 4 bp overhangs. You can then use the Klenow fragment...Science 239 (1988): 487-491. PubMed PMID: 2448875. 4. Xia, Yongzhen, et al. "New insights into the QuikChangeTM... Brush up on Your Cloning Techniques Learn More about Plasmids in Our Plasmids 101 Series Use CRISPR to...

...with the crRNA, (3) three transposase proteins, and (4) the donor DNA. By using a multi-spacer CRISPR array...Using an Engineered CRISPR/Cas System. ACS Synth Biol 4:723–728 . https://doi.org/10.1021/sb500351f Dong C...diverse bacteria with Mobile-CRISPRi. Nat Microbiol 4:244–250 . https://doi.org/10.1038/s41564-018-0327-...CRISPR Featured Topic Page Learn more about Cpf1 Find out about CRISPR Multiplexing Resources on Addgene.org...coli and other bacterial species. Read on to learn about the tools available for bacteria and some of the...that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab published a ...as a collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases...

...scheduled for March 4th from 2:30-4:00 pm ET.  You can learn more about the report's recommendations and...

... and you’re venturing into the uncertain terrain. 4. Never trust the first 20-30 bases of a DNA sequencing...and troubleshooting your DNA sequencing results Above (top right corner of this post) is an example of...

... translational. Ex: the discovery of GFP. Chapter 4: Goals of the Center for Regenerative Meidince - Understanding...promote scientific sharing. Listen to learn more about Darrell, his research, and his focus on open science...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Podcast.     Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids...Plasmids from the Kotton Lab Learn more about Stem Cells ...

...mechanisms that cause plasmids to be incompatible. 4. RNA Extraction Without A Kit by Leah Schiesow We... highlighted reagent repositories and other collaborative efforts in the scientific community to speed...store my plasmid in TE, Tris, or water? And what about temperature? Find the details in this blog post....dominates DNA quantification discussions, but what about the other methods to quantify DNA? Here we cover...Incompatibility by Leah Schiesow If you're thinking about hosting multiple plasmids in one cell, it might ...

...SSC-H / SSC-A gate would work just as fine (Figure 4). As H and A scale in proportion (while H and W do...                                                                                                         B.    Figure 4: (A) Using the proportional relationship between FSC-H...lymphocytes in a mouse tumor sample You made it! So much about the theoretical background. Let’s dive into the ...often choose a looser gate here. I am not worried about including irrelevant events in my analysis, as these...population, acquired from the previous gating pipeline, about 53% of my cells are dendritic cells and ~13% are...ve covered in this blog post. First, you learned about the types of scattering that occur when a cell is...

...fluorescent under acidic conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP ... biology 27 (2015): 39-45. PubMed PMID: 26087123. 4. Kumagai, Akiko, et al. "A bilirubin-inducible fluorescent...deleting residues. Only about 10 amino acids can be deleted from the C-terminus and about 5 from the N-terminus...cylinder with a length of 4.2 nm and a diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel...based on cofactor binding proteins as described above. The smallest tags are peptides that are engineered...

...to open scientific research data around the world. 4)      The Annals of Internal Medicine published systematic...study probably doesn’t say as much about open access as it does about the state of peer review. 8)     ...University Open Data Access (YODA) Project, “a collaborative endeavor guided by aspirations to promote open...

...targeting Golgi, ER, and endosome membranes) (Figure 4). The authors used SuperPLD to manipulate phosphatidic...fluorescent labeling via click chemistry.    Figure 4: SuperPLD hydrolyzes membrane PC to PA. A) OptoPLD... Check out our Optogenetics Guide to learn more about how these modules work, and read on to learn how...to the cell membrane isn’t powerful enough, what about altering the chemical composition of the membrane...

...topics from the Science by the Pint series. Chapter 4: The SITN Podcast, Sit'N Listen - In their podcast...To learn more about the STIN's podcast, email [email protected]    For more info about starting your...evens where scientists can grab a beer and chat about their work at a local bar. It doesn't have to be...1: Introduction - Vini and Amy tell us a little about themselves. Chapter 2: What is Science in the News...science communicators. In this chapter we learn about some of their events - science seminars, science...the lab and working with SITN. She points to collaboration and delegating as keys achieving success. Chapter...your own SciComm organization and to learn about some of SITN's other initiatives, email [email protected]...

...: mouse, fruit fly, yeast, zebrafish, and worm. 4. Early Career Researcher Toolbox: Free Tools for Making...mNeonGreen, mTFP1, and mWasabi by Jennifer Tsang Learn about mNeonGreen, mTFP1, and mWasabi and find plasmids...Broccoli, Spinach, and Corn, for example. Learn more about the aptamer soup in this blog post. 5. Five Popular...

...names of everyone at Addgene, and send random sets of 4 people out to lunch.   I think these random lunches...for the company, but also talking about sports, movies, and just about anything. As our organization grew... with our western blots. Thoughts about sharing Addgene is all about helping scientists share between ...culture of openness and trust that comes from caring about your colleagues on a personal level, and employees...often cite lunch as one of their favorite parts about working at Addgene. We’re a small non-profit, so...indication of the stiff competition or says something about my cooking.   This is a creative (and delicious)...for people in different labs to chat informally about their challenges, successes, and expertise. Who ...

...The Addgene-Seeding Labs plasmid grant is a collaborative endeavor providing plasmids to researchers in... of Zimbabwe, Zimbabwe Dr. Idah Sithole-Niang (above, left) is an Full Professor in Biochemistry with... CRISPR effect observed in bacteria. Learn more about the different CRISPR applications in plants.     ...Rodrigo Arturo Rivera Solís, Associate Professor (above, far right), thrives to make affordable biosensors...detect glyphosate via H2O2, one of the reaction’s metabolites. One useful genetically encoded H2O2 probe is... to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees, all who...Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant Check Out...

...Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas...expression of cDNAs and shRNAs with a reporter gene, or (4) multiple sgRNAs with Cas9 for genome engineering....Sladitschek HL and Neveu PA. PLoS One. 2015. Apr 24;10(4):e0124958. LOC8-ing mitochondrial fusion with a...system was also designed for a short screening time (<4 hours) in order to avoid complicating effects from...improving the strength of the promoters used; and 4) optimizing expression of both the light sensor and...The proteins encoded in this set range in size from 4 to 156 kDa, enabling the user to characterise the ...new plasmid tool from the past year? Let us know about it in the comments or shoot us an e-mail at blog...

...facing towards or away from each other.  Figure 4: How to design a conditional allele with Easi-CRISPR...Biol. 18 (2017) (1):92. DOI: 10.1186/s13059-017-1220-4 PubMed PMID: 28511701 PubMed Central PMCID: PMC5434640... on CRISPR Mouse Models Learn about Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) ... a variety of species, including the standard laboratory mouse. It’s also made targeted insertions relatively...process of conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult... a mouse embryo to create chimeric mice. This laborious process takes ~8-10 months. With CRISPR (B), you...directly into a zygote and obtain a knockout mouse in about 3 months. Image courtesy of Wenning Qin. Single-stranded...

... Jan;103(1):303-313. doi: 10.1007/s00253-018-9485-4. Epub 2018 Nov 3. PMID: 30392122. Pfeiffer, Barret... system in Caenorhabditis elegans.” Nat Methods 9(4) (2012): 391-5. PubMed PMID: 22406855. PubMed Central...can be used in the repressible manner described above, as well as the inducible manner detailed in our...necessary to have all 3 of the different binary systems above if they all behave very similarly?” The simple answer... at the same time.  The 3 binary systems listed above are orthogonal, meaning that they do not affect ...Srikrishnan. “Introduction and expression of genes for metabolic engineering applications in Saccharomyces cerevisiae...