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Showing: 501 - 520 of 951 results
  1. 10 Basic tips for mammalian cell culture

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    Blog Post
    ... 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 ...
  2. AAV Vector Quality Control: Going the Extra Mile with NGS

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    Blog Post
    ...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...
  3. Tips for a 1st time AAV user (by a Rookie AAV user)

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    Blog Post
    ...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...
  4. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

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    Blog Post
    ...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...
  5. Plasmids 101: Protein tags

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    Blog Post
    ...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...
  6. Hot Plasmids Spring 2024

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    Blog Post
    ...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...
  7. Plasmids 101: Broad Host Range Plasmids

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    Blog Post
    ...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...
  8. Hot Plasmids - November 2023

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    Blog Post
    ...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...
  9. Viral Vectors 101: Preparing Pooled Libraries

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    Blog Post
    ...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...
  10. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

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    Blog Post
    ...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...
  11. CRISPR-based Genome Editing Systems For Editing in E. coli

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    Blog Post
    ...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...
  12. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

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    Blog Post
    ...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...
  13. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    Blog Post
    ...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...
  14. Career Insights: Technical Support Specialist

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    Blog Post
    ...expertise with that of the company. To increase your chances at successfully applying for a position, you should... in a technical support role. What general characteristics do jobs in technical support have? There are... less time consuming and/or less technically challenging. Alongside there is a second-line team that takes... ways to communicate with customers. Online live chat is a convenient alternative for asking a quick question...
  15. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...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...
  16. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

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    Blog Post
    ...for researchers looking to generate long-lasting changes in cells. However, for some applications, integration...numerous variants have been developed with different mechanisms to inactivate the integrase.  The IDLV life cycle...construct has the D64V mutation, a single amino acid change (from aspartic acid to valine at position 64) which...Kempski, H., Brugman, M. H., Pike-Overzet, K., Chatters, S. J., de Ridder, D., Gilmour, K. C., Adams, ...
  17. Replacing Paper: Tips for Choosing an Electronic lab Notebook

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    Blog Post
    ...How are you currently exchanging information with your team? If you need to exchange information regarding...which they are sure that there will be no sharp changes in costs once they establish the system within ...consolidated to one place, easily retrievable, searchable and traceable. However, switching from paper ...
  18. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

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    Blog Post
    ... of reasons to consider using genome editing to change the genetic code of plants, including the development... system in most organisms, some plant-specific changes to CRISPR components are necessary to enable genome...try your hand at CRISPR. References Belhaj K, Chaparro-Garcia A, Kamoun S, Nekrasov V (2013) Plant genome.../mmbr.67.1.16-37.2003 Khatodia S, Bhatotia K, Passricha N, Khurana SMP, Tuteja N (2016) The CRISPR/Cas...
  19. How to Negotiate a Successful Lab Start Up Budget

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    Blog Post
    ... do you need [for your lab start up]?” A hiring chair or dean will often ask a faculty candidate. Sweaty...focusing on long term investments. For example, purchasing “used” equipment can save you money in the short...their tangible resources. For example, if a dean/chair asks “how much do you need to start your lab”. You..., working with this question is second nature. Channel your inner curiosity and use your research training...
  20. Plasmids 101: Dimers and Multimers

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    Blog Post
    ...giving greater concentration of cut DNA without changing the size. As a result, a restriction digest of...reducing multimerization and define recA-independent mechanisms. Avoid overgrowing the bacterial culture Another...PMCID: PMC3060453. Bazaral M, Helinski DR. Characterization of multiple circular DNA forms of colicinogenic...Adv. 2009 Jul-Aug;27(4):353-70. doi: 10.1016/j.biotechadv.2009.02.003. Epub 2009 Feb 20. PMID: 19233255...
Showing: 501 - 520 of 951 results