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We narrowed to 993 results for: Des

Showing: 401 - 420 of 993 results

  1. Fluorescent Proteins 101: Luciferases

    Type
    Blog Post
    ...abdomen to produce yellow-green light (Figure 2). Despite the title of this post, this reaction is not fluorescence...fluorescence! Chemiluminescence, which includes all bioluminescence, is the production of light by a chemical...assays (low background, no photobleaching) but provides flexibility by pairing the luciferase with a variety...luciferase output at different wavelengths. Addgene provides empty backbones with the luciferase gene, a variety...

  2. Viral Vectors 101: The AAV Retrograde Serotype

    Type
    Blog Post
    ...trillion connections (synapses.)  In this post we’ll describe the AAV retrograde serotype (AAVrg) and how it... this can cause challenges for the experimental design as these vectors can exhibit spurious expression...efficiency can have a major impact on experimental design and interpretation of results. Take extra care ...References 1. Tervo, D. Gowanlock R., et al. "A designer AAV variant permits efficient retrograde access...

  3. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    ...but can be minimized with high powered research designs. Unanticipated discoveries Finally, there are good...rates, should be reported as a new discovery that deserves confirmation with new data and a new, preregistered...Guidelines; preregistration to increase clarity in study design and analysis; removing publication bias with Registered...Science, Dr. Mellor worked with citizen scientists to design and implement authentic scientific research and...

  4. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ... NEB's Resource describing Dam and Dcm Methylases Discovery of DpnI Review describing dam-/dcm- E. coli...mutagenesis. During this process, incorporation of a desired mutation into your plasmid of interest by PCR generates...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which...

  5. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...cleaved precisely as predicted with no addition of nucleotides to the 5’ spacers. The 3’ ends were modified ...advantage over other methods that favor certain nucleotides at the 5’ end. PTG construct schematics. Scissors...the plasmids made by Xie et. al are specifically designed for plant expression, PTGs will likely be adaptable...worries, I've summarized the three strategies described in this post in the graphic below - including ...

  6. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ... for over 10 years. Many successful scientists describe having a “posse” of mentors as one key to their...Ellen Daniell. In “Every Other Thursday,” Daniell describes a group of 7 women scientists who met every other...increases the diversity of input and perspectives Provides a good mechanism for accountability that makes...leadership and communication skills. The leader also provides materials such as pre-reads, videos or exercises...

  7. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...shown in the figure below. Their most successful design utilized Magnet photoswitchable proteins derived...architecture. Yu et al., 2020 FAST BphS from R. sphaeroides Split Cas9 Genome editing by NHEJ and HDR. Can...Cas9 Richter et al., 2016 tsRC9 RsLOV from R. sphaeroides Cas9, dCas9 Gene editing Zhou et al., 2018 ...al., 2016 CRISPR-plus Photocleavable ssDNA oligonucleotides Cas9, photocaged-gRNA Gene editing Manna...NPPOC-caged trimethoprim (TMP) Cas9 fused to destabilizing domains (Cas9-DHFR) Gene editing, transcription...

  8. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ... as the OG (as the kids say) affinity reagents deserved to be celebrated, there are many other amazing...its thorns and every affinity reagent has its downsides. These reduced-in-size options often have reduced...°C and can refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically...a wide variety of targets, including proteins, peptides, amino acids, drugs, metal ions, and cells. They...

  9. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    ...molecular biology experiments was in the job description, but this skill is not just about reacting to...puzzling through your broader research questions and designing future experiments. Being able to work through...research non-academic careers, I always felt that descriptions of transferable skills seemed fuzzy or ill-defined...most likely cause, you implement an experiment designed to fix the problem. Then you assess whether your...

  10. Antibodies 101: Conjugation

    Type
    Blog Post
    ...we would be remiss not to at least mention. Oligonucleotides allow for sensitive detection of low abundance...strong protein-protein interactions with other peptides, like an Fc-binding domain (Figure 1).  ...Strong protein-protein interactions with other peptides. Created with BioRender.com.     Many biologists... conjugation. But hopefully, this introduction provides a good base from which to dive deeper!References...

  11. The time and cost required to make a plasmid

    Type
    Blog Post
    ...in science – the time spent by the scientists designing and performing the experiments. This cost varies...quickly with the postdoc who’s really good at primer design – but first, you have to know what resources exist...well) plate! How much do repositories help?   Designing precise calculations for any of the numbers in...engagement with and use of repositories like Addgene provides large-scale savings. How large? Using Doench’s...

  12. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...and SMASh degron tagging vector! Destabilizing domains Destabilizing domains (DD) are akin to degron ...   Fig. 2 Tetracycline on/off promoters and modes of action Check out Addgene's Lentiviral Tet-on ...post-translational level, unlike most of the other mechanisms described. Fig. 3 dTag mechanism of protein degradation...

  13. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ... plasmid maintenance during cell division. ccdB codes for the toxic protein (CcdB) that acts as a DNA ...death. ccdA, another gene found in the ccd operon, codes for the antitoxin protein (CcdA) that protects the...of ccdB, leading to positive identification of desired clones.   ...Becomes a powerful cloning tool... efficiently, as those that do not contain the desired insert should not grow.   Figure 2: Image...

  14. Developing Lab Management Software for Biology

    Type
    Blog Post
    ...another. If you are using barcodes (and you should be!) bear in mind that your barcodes are being applied to...containers, not to the contents. Tightly coupling barcodes and containers might not be such a bad idea, tightly...Resources on the Addgene Blog Learn about How We Use Barcodes in the Lab Get Tips on How to Keep a Lab Notebook...

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

    Type
    Blog Post
    ... nuclease activity of the Cas9 nuclease itself. Design and testing of eSpCas9 Looking at the structure...and the negatively charged DNA and therefore destabilize basal strand separation. This decreased strand... choice and get improved on-target specificity. Design and testing of SpCas9-HF1 Kleinstiver et al and...Joining Repairs Double Strand Breaks Get Tips on Designing Your gRNA Browse Our CRISPR 101 Series Resources...

  16. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ... allele decreases. Crossing mice to generate a desired phenotype Instead, genetic engineering should be...in the journey towards producing mice with your desired combination of mutations. Assuming your parental...generate mice that are homozygous for all of your desired genes. Remember Punnett squares from high school... are homozygous (or hemizygous) at all of your desired alleles to conduct your experiments. You should...

  17. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...newer generation lentiviral packaging systems were designed to prevent the formation of replication competent...use 3 plasmids; an envelope plasmid that usually encodes VSV-G, a packaging plasmid containing structural...transfer plasmids with small inserts (such as CRISPR guides) while a single 72h harvest is best for larger ...the best titer possible. While this information provides a great starting point, it may not translate to...

  18. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ...soCoChR-expressing system. Taken together, Shemesh et. al. describe a novel system for the precise mapping of neuronal...the single-cell level. Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid...information about spatial and temporal organization. The design use to detect PKA activity can be generalized to...choice for FRET assays where a dark acceptor is desired due to its very low quantum efficiency. ShadowY...

  19. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...tools, they do have undesired off-target editing effects. ABEs have been described to introduce base changes...efficient. Informed by these mutations and previously described TadA mutations (Gaudelli et al., 2017), they modified...heterodimeric fusion of wild-type TadA and TadA7.10, they designed one set of ABE8’s similarly (ABE8.x-d). In order...

  20. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    ... developed based on 100 structure-guided spike designs and has improved protein yields and stability. ...Plasmids deposited from Alejandro Balazs’s lab includes those for generating pseudovirus and those encoding...Liu Y, Chou C-W, Byrne PO, Hjorth CK, Johnson NV, Ludes-Meyers J, Nguyen AW, Park J, Wang N, Amengor D, ...Finkelstein IJ, McLellan JS (2020) Structure-based design of prefusion-stabilized SARS-CoV-2 spikes. Science...
Showing: 401 - 420 of 993 results