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We narrowed to 806 results for: cha;

Showing: 341 - 360 of 806 results

  1. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise the cost anywhere...one-off projects, or even save yourself hours by chatting quickly with the postdoc who’s really good at ...harder it is to throw in last minute experiments or changes. Fun fact: Addgene’s high-throughput approach means...

  2. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    Published
    May 11, 2021, 1:15 p.m.
    ...monkeys. Surprisingly, we found that this small change made a big difference. Replacing the mCherry with...does highlight the importance of a careful characterization of transgene expression. Which tag should ...RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method...Raper J, Hu X, Paré J, Bonaventura J, Richie CT, Michaelides M, Mueller SAL, Roseboom PH, Oler JA, Kalin NH...

  3. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...CRISPR/Cas9 genome editing can result in unwanted changes at non-target sites. Means to decrease these so-called...between the non-target strand and a positively charged groove formed by the Cas9 HNH and RuvC nuclease...al reasoned that, if they decreased the positive charge in the HNH/RuvC groove, then they could weaken ...interactions between the groove and the negatively charged DNA and therefore destabilize basal strand separation...

  4. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    Published
    April 22, 2015, 2:06 p.m.
    ... with Cas9 guided by different promoters. The challenge in building genetic circuits is getting enough...shape of the response function or how the output changes as a function of input. Usually that function is...enough genetic parts for complexity. We have characterized almost 600 terminators. That gives us enough... we’ve really been making tons of parts and characterizing the software to help you put it together. The...

  5. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...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...

  6. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...light-activated ion channels. Similarly, chemogenetic tools are designed channels that are chemically ...introduced in neurons and depending on the choice of channel protein, they can trigger neuronal excitation or...adeno-associated virus. Curr Protoc Microbiol. (2013); Chapter 14:Unit14D.5. PubMed PMID: 23686825. PubMed Central...

  7. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...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...

  8. How to Prepare for an Industry Interview

    Type
    Blog Post
    Published
    July 9, 2020, 1:15 p.m.
    ...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...

  9. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ...Shyu YJ, Liu H, Deng X, Hu CD. Biotechniques. 2006 Chang-Deng Hu Green sfGFP Split super-folder GFP, our...protein-protein interactions in vivo. Blakeley BD, Chapman AM, McNaughton BR. Mol Biosyst. 2012 Brian McNaughton...Shyu YJ, Liu H, Deng X, Hu CD. Biotechniques. 2006 Chang-Deng Hu Venus Improved N-terminal fragment VN155...signal-to-noise ratio. Kodama Y, Hu CD. Biotechniques. 2010 Chang-Deng Hu mVenus pET-BiFC contains both fragments...

  10. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...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 ...

  11. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    Published
    Jan. 21, 2015, 1:30 p.m.
    ...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...

  12. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ... 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 ...

  13. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...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...

  14. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...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...

  15. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...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...

  16. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    Published
    May 22, 2015, 4:58 p.m.
    ...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...

  17. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.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...

  18. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...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...

  20. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...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...
Showing: 341 - 360 of 806 results