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We narrowed to 845 results for: ins

Showing: 541 - 560 of 845 results

  1. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ... Rho-independent termination is also known as intrinsic termination, and it relies on the formation of...tail targets the RNA to the degradosome, which contains enzymes that cut RNA not protected by secondary...complex and involves two RNA polymerase-associated proteins, CPSF and CstF, which are responsible for recruiting...Cell. 1987. PMID: 3568131.  Hager S, Frame FM, Collins AT, Burns JE, Maitland NJ. An Internal Polyadenylation...

  2. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    Published
    Aug. 9, 2022, 1:15 p.m.
    ...unreliability: cross-reactivity (binds to non-target proteins), variability (separate batches can perform differently...mostly uses whole-mount IHC techniques on rodent brains and is expanding to more complex brain systems ...larger-volume antibodies for thicker tissues and whole brains, we chose monoclonal antibodies (mAbs) and their...negative control which does not have primary and only contains secondary antibodies, allowing us to only image...

  3. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...indicators (GECIs) are fluorescent proteins linked to calcium binding proteins (e.g., calmodulin or troponin... changes with Ca while another wavelength is insensitive. Additional red sensors based on mRuby (jRCaMP1...Addgene.org Addgene’s AAV Guide Fluorescent Proteins: Biosensors Addgene’s Viral Services  Resources...

  4. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ... Khan Khilji, a PhD student at the Max Planck Institute of Colloids and Interfaces. Here are some tips...split cells Don’t be lazy about splitting cells, instead try to form a routine. Twice a week often works...take a vial of the ‘mint condition’ master stock instead of ‘borrowing’ cells that have already been passaged...soon be joining her PhD lab at the Max Planck Institute of Colloids and Interfaces. Sana is particularly...

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

    Type
    Blog Post
    Published
    Sept. 12, 2017, 1:44 p.m.
    ...helper plasmids or the cell genome can be packaged inside the vectors during production. These impurities...removed during the purification step because they are inside the virus itself. While these impurities are generally...NGS at Addgene to sequence all the DNA packaged inside the viral particles (Figure 1). Briefly, packaged...to the contaminant. Each contig is then BLASTed against the NCBI nucleotide collection database, and it...

  6. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...November 18, 2016): Researchers from a variety of institutions recently reported their inability to recapitulate... and development before it can truly be pitted against CRISPR head-to-head.  Exciting features of NgAgo...and fussy to optimize. One reason could be the instability of the supposedly alien 5’-phosphorylated ssDNA...Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo: Genome-editing by a ...

  7. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...these therapeutics. Jesse Bloom’s lab at the Fred Hutchinson Cancer Research Center developed a pooled plasmid...created a novel nanocompartment for transporting proteins in yeast by using murine polyomavirus virus-like...VLPs ). This artificial nanoparticle can protect proteins from unfavorable interactions in the cell, improving...zCas9i cloning kit is a plant cloning kit that contains a series of vectors and DNA modules for assembly...

  8. PhD Applications After COVID

    Type
    Blog Post
    Published
    Jan. 5, 2023, 2:15 p.m.
    ...department during the application process. Many institutions recommend that you reach out to potential advisors...you can’t visit. By talking with people at the institution, you can get a better sense of the environment...answer process can also help you to see if the institution is right for you outside of just the program ...options are very handy in moments like that! For instance, during my application cycle, two schools on opposite...

  9. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...antimicrobials could become our newest line of defense against bacteria. Created with BioRender.com. ...resistance using multiple approaches. Major restaurant chains including Chipotle and Panera Bread have pledged... just the method needed to jumpstart the fight against antibiotic resistance. Advantages of CRISPR over...strategy can help us win back ground in the fight against bacterial superbugs. References 1. Citorik, Robert...

  10. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...D-F)  High-magnification images immuno-stained against dopamine beta-hydroxylase (DBH, magenta) to label...and that it often provides incredibly valuable insight into one’s work. As a community, we must therefore...remove the stigma around sharing negative data and instead focus on getting this information out into the ... A., Hsu, R., Kalwani, L., Sierra-Mercado, D., Robinson, J. O., Outram, S., Koenig, B. A., Pereira, S....

  11. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...labs of David Root and John Doench at the Broad Institute have developed human genome-wide CRISPRko (knock...CRISPRi libraries utilize an engineered Cas9, which contains multiple point mutations in the nuclease domain...potential off-target effects. The Dolcetto library contains two sets of sgRNAs; A, the top 3 ranked sgRNAs... sequences can also be modified to include RNA domains that recruit additional transcription factors. ...

  12. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    Published
    July 30, 2024, 1:15 p.m.
    ...process that isn’t as simple as it seems. Take, for instance, the GCaMP-encoding viral vectors available in...service. While they were both requested, GCaMP6 remains the more requested option, and jGCaMP8 appears ...compared to other AAVs preps from 2016–2023. B) Inset showing distribution of only GCaMP6, jGCaMP7, and...our own repository data, giving us significant insight into which vectors are likely to be of most use...

  13. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...medical journal. Decades later and after years of painstaking work trying to isolate the causative gene for...recently used the CRISPR/Cas9 system to both precisely insert the MECP2-R270X mutation into the MECP2 gene in...fine-tuning dosage so that enough wild-type MECP2 is made inside the cells, but this is exciting progress in tackling...:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, Lichtarge O, Zoghbi HY (2013) An AT-Hook...

  14. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    Published
    Nov. 12, 2020, 2:15 p.m.
    ...growing polypeptide chain. One part of the tRNA contains the anticodon,  a three nucleotide sequence complementary...become extremely useful in expressing functional proteins in hosts that do not naturally express that gene...host you are using or come from an organism that contains expression limiting regulatory elements or use...10.1038/nsmb1031 Gustafsson C, Govindarajan S, Minshull J (2004) Codon bias and heterologous protein expression...

  15. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    Published
    March 31, 2020, 1:15 p.m.
    ...with the replicon- the part of the plasmid that contains the origin of replication and the replication ...inhibiting translation of replication machinery proteins, or directly inhibit replication by binding to...walking” the same plasmids to one daughter cell, instead of partitioning each type of plasmid into a separate...2015) Imaging centromere-based incompatibilities: Insights into the mechanism of incompatibility mediated...

  16. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    Published
    Jan. 17, 2019, 1:38 p.m.
    ...reproducibility is limited in two ways – only data inside the ELN is reproducible, and only the highly restricted...Goettingen, and several other large research institutions. The following graphic depicts the ways in which...banks and sequencing files is likely stored in institutional and lab servers specifically designed to hold...illustrated in this video, and to data stored on institutional and lab servers. Connectivity to other tools...

  17. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...Transition mutation (C>T, G>A, A>G, and T>C) installation by base editors. Pathways that compete with ...developed plant adenine base editor (ABE) vectors to install A>G or T>C mutations in the plant genome. We constructed...probably due to the presence of nucleosome or other proteins. You can detect base editing as early as 48-72...Ag-biotechnologist in the National Rice Research Institute, Cuttack, Odisha, India. Currently Kutubuddin ...

  18. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...purified viral vector solution (which contains viral capsid proteins) as opposed to purified DNA. This may...see below), since each viral particle typically contains one viral genome. Other than a theoretical maximum...mitigated by using purified viral DNA as the template instead of the intact AAV particle. Finally, the precision...

  19. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ...every experiment, an investigator needs a standard against which the results can be compared; results from...single experiment wherein Plasmid A (encoding shRNA against human Gene X in backbone Y) was transfected into...included as these provide the baseline/standard against which other samples can be compared. The Empty ...For example, in the experiment above, Plasmid A contains an shRNA that targets human Gene X and human cells...

  20. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...within living cells. By coupling fluorophores to proteins, pioneers of this technique were able to directly...FRET-based biosensors, and interactions between DNA and proteins. FRET, in theory, is an easy technique, but it...to studying an unknown interaction between two proteins, these parameters are difficult to overcome but...FRET_teaching_module.pdf Day RN, Davidson MW. Fluorescent proteins for FRET microscopy: monitoring protein interactions...
Showing: 541 - 560 of 845 results