...of the theme of my interview stories as the characteristic which the story demonstrates I possess.   Bullet...to present about your prior research. This is a chance to highlight your prior accomplishments and demonstrate...carefully. And even if you don’t have technical challenges, your interviewers might.  You’ll probably still...their hand to ask a question or type it into the chat. Either way, just let the interviewers know up front...

...Indicate the amino acid change as well as any functional consequences of that change. For example, “Mutated... Answer “What does this plasmid do?” in <200 characters. A descriptive purpose focuses on the practical...Addgene Blog Find Articles Citing Addgene Plasmids Searchable and Sortable gRNAs for Your Next CRISPR Experiment...

...experiments are either not accessible or too challenging for amateur scientists. Planarians, wormy creatures...cell types and for viewing how gene expression changes through time or after genetic manipulation. The...tissue sample, several washes, and temperature changes to visualize the expression of specific genes. ...genes of interest and was first discovered and characterized in an unrelated worm (C. elegans) [2]. Delivering...

...tasks, both of which are factors in the technical challenges they can present. Many researchers find themselves... following transfection. In other words, when exchanging the medium after transfecting cells, replace ...electrostatic forces between these two negatively-charged membranes (Davis et al., 2002). Polybrene is one...306. PubMed PMID: 10634178. 8. Murnane, John P., Michael J. Yezzi, and Barbara R. Young. "Recombination ...

...the only right way, and Indecisives, who keep changing their minds and have trouble making decisions.... of your supervisor’s management style, bring challenges to his or her attention as soon as you can. It... action items. This will give your supervisor a chance to review what you have discussed and add suggestions...mutually beneficial solutions. If you package your challenges with your supervisor into learning opportunities...

...titration by qPCR.   Pro tip! An optional media exchange can be performed 12–18 hours post-transfection...the density gradient material by dialysis/buffer exchange and further concentrate the virions if needed....Lock, M., McGorray, S., Auricchio, A., Ayuso, E., Beecham, E. J., Blouin-Tavel, V., Bosch, F., Bose, M., ...., Francis, J. D., … Snyder, R. O. (2010). Characterization of a recombinant adeno-associated virus type...

...genomes, cloning vectors, and helper plasmids (Chadeuf et al., 2005, Wright et al., 2011). In fact, the... software. References and resources References Chadeuf G, Ciron C, Moullier P, Salvetti A (2005) Evidence...Redon R, Moullier P, Léger A (2015) Advanced Characterization of DNA Molecules in rAAV Vector Preparations....2015.32 Wright JF, Zelenaia O (2011) Vector Characterization Methods for Quality Control Testing of Recombinant...

...AAV-containing 40% fraction is collected and buffer exchanged to remove the iodixanol and concentrate the purified...or the virus can be stored at 4 °C and buffer exchanged the next day. Refer to the AAV Iodixanol Gradient...correctly. When aliquoting purified and buffer exchanged virus, remember to make one or two small aliquots...vivo use. Current protocols in molecular biology, Chapter 16, Unit 16.25. PubMed PMID: 18265393. Additional...

... 37 °C and 5% CO2 and get agitated with sudden changes in temperature (especially if you have air conditioning... to recover after they are passaged. This could change their growth rates and morphology, rendering their...numbers Keep an eye on the passage number. Cells change their growth rate and morphology at high passage...Additional Resources on the Addgene Blog The Challenges of Cell Culture Cloning Mammalian Cells with ...

...through wild populations. Whereas most genomic changes impose a fitness cost and are eliminated by natural...previous gene drive and thereby undo the genetic changes driven by the earlier intervention2. While an initial.... PubMed. Gantz, V. M. & Bier, E. The mutagenic chain reaction: A method for converting heterozygous to...10.1126/science.aaa5945. PubMed. DiCarlo, J. E., Chavez, A., Dietz, S. L., Esvelt, K. M. & Church, G. M...

...purify proteins can face any number of technical challenges depending on their protein of interest. After...After several decades of trying to address these challenges, researchers have amassed a considerable molecular...for your protein. If you’re working with an uncharacterized protein, or a protein for which a good antibody...KETAAAKFERQHMDS  1.8 Solubility and Affinity Abundance of charged and polar residues improves solubility; good for...

...protected by Cas9. This is similar to the proposed mechanism for engineered pegRNAs (epegRNAs), which add a...of mouse monoclonal antibodies generated and characterized by the Geiss Lab in their effort to fill a void...cells. Plus, Voltron2’s fluorescence is tunable by changing the dye that becomes conjugated with its HaloTag...gene expression regulation, enhancing both the mechanisms of delivery and the potential for therapeutic...

...Design). This modular approach combines an interchangeable set of fusogenic and targeting components to...where the fusion and targeting components are interchangeable. Image from Strebinger et al., 2023, reused...local proteome, but some applications have been challenging or inaccessible. The Ting lab recently made two...They demonstrated LOV-Turbo in neurons, in pulse-chase experiments, and more. Find LOV-Turbo plasmids here...

...the chances of a cell being infected by only one viral particle, but it also increases the chances that...presentation deceive you; pooled libraries can be quite challenging to use! In this post, I’ll walk you through how...virus. Let’s start there!     Figure 1: Flowchart showing the process for using pooled lentiviral...

...General Meeting seven other times I had subsequently changed fields and my cohort of friends and colleagues ...evolutionary lineages share genes that can improve chances of survival for an individual, and eventually, ... cells regulate gene expression in response to changes in cell density. At the mixer, they meet other ...Dill-McFarland, Lisa Gorski, Mark O. Martin, Maureen Berg, Michael D. L. Johnson, Tiffany Lowe-Power, Eric Griffis...

...incentivize good mentorship: AMA mechanics: Be prepared! The mechanics of doing the AMA were interesting...Getting on a good science career path can be challenging these days, but for many of us, scientist is ...Science through Video with the Addgene Youtube Channel Check Out Vidoes of Gender & Science from LabLinks...

...effects. The flaws of RNAi screens are well characterized and strategies exist to control for these faults...also reproduced several previously observed characteristics that influence gRNA off-target activity: 1)...23394947. Pubmed Central PMCID: 4299491. 5. Bassik, Michael C., et al. "A systematic mammalian genetic interaction...available from Addgene Other Resources Watch Dr. Michael Bassik’s lecture on “Multiplexing with CRISPR Screens...

...Understanding the characteristics of natural plasmids and how they function in nature can inform us on...often have a restriction barrier as a defense mechanism, and they can degrade foreign bacteria through...expressed. For example, scientists can incorporate interchangeable antibiotic resistance cassettes (Prior et al...Schmidhauser, T. J., Ditta, G., & Helinski, D. R. (1988). Chapter 15—Broad-Host-Range Plasmid Cloning Vectors for...

...following changes. In pCas9, we 1) replaced the promoter of gRNA-pMB1 with PrhaB, 2) changed the replicon...same year. Her research is focused on the escape mechanism of genome editing in microorganisms which could...cells to understand the genetic and biochemical mechanisms behind their distinct phenotypes and to design...

...of reprogramming (or “Yamanaka”) factors, they changed the landscape of regenerative medicine. Their work...viral vectors are preferred. This is to avoid the chances of insertional mutagenesis, which is the integration...replacing C-MYC with L-MYC [10, 11]. While the mechanisms underlying the enhancement of iPSC generation...Induced Pluripotent Stem Cells (iPSCs): Benefits, Challenges and the Road Ahead." Journal of Alzheimer's disease...