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We narrowed to 66 results for: ha/1000

Showing: 41 - 60 of 66 results
  1. General Transfection

    Type
    Protocol
    ...ensure that it has not drifted. Filter the solution through a 0.22 µm membrane. Aliquot 500–1000 µL into sterile...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...
  2. Typing CRISPR Systems

    Type
    Blog Post
    ...these adaptive bacterial and archaeal immune systems has led to extreme sequence divergence that requires ...landmark publication in 2013. Since that point, it has become the most engineered Cas enzyme, with adaptations... CRISPR type that exclusively targets RNA. Cas13 has become a natural successor to traditional RNA interference...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 Additional resources on ...
  3. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...effective than both AAV9 and AAV5, which has tropism for lung cells, and has increased overall transduction throughout...this definition: A systemic capsid is a capsid that has been engineered to have enhanced function after intravenous...both). Therefore, a high-efficiency systemic capsid has better efficacy than AAV9 and/or its intermediate...used in LY6A-nonpermissive mouse strains. **AAV1-X1 has not been tested in rats. For more information on ...neurons in the CNS (Chan et al., 2017). AAV-PHP.eB has frequently been used for screening candidate regulatory...depositing manuscript) across the brain. AAV-BI-28 has been tested in C57BL/6J mice and effectively transduces.... 4c, Jang et al., 2023; Brown et al., 2021.) It has a similar transduction rate to AAV-PHP.eB in the ...
  4. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 20–200 µL multichannel...Microcentrifuge tubes, Neptune 3745.X Pipette tips, 1000 µL, VWR 76322-154 Pipette tips, 10 µL, VWR 76322...periodically to determine when the desired color change has occurred. The color will change from yellow to blue...experimental conditions. When the desired color change has occurred, gently remove the plate seal. Using a multichannel...
  5. The Future of Research Symposium Boston 2015

    Type
    Blog Post
    ...Research will be held from 22-24 October. This blog has been contributed by guest blogger and Future of Research... of a system that, despite all of its successes, has many deep-rooted flaws (1). As we discovered through...example, Harvard’s Therapeutics Graduate Program has a compulsory internship to be taken in an industry...Marder, professor of Biology at Brandeis University, has recently published an article arguing against reducing...Labor and Worklife Program at Harvard Law School. He has continually promoted the cause of increasing data...Research: a perspective from junior scientists." F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central...
  6. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...CRISPR/Cas9 technology has been a breakthrough for the genome-editing field. It has the possibility to treat... applications.” Li’s team noted that CRISPR/Cas9 has been used to generate mutations in a wide range of... expanded organoids. “CRISPR in adult stem cells has huge potential, because the genomes of adult stem...& Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6. Wu et al. 2020. . “Advances in CRISPR/Cas-based...
  7. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by The RNAi Consortium, also known as the...Plasmid Kit Retroviral Barcoding Library (Kit # 1000000097) The Winslow lab barcoding library consists of...Multiple Lentiviral Expression System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral Expression) system...
  8. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ... in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes...protein interactions in living cells. Since then FRET has also been used to measure conformational changes,...brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP. The stoichiometry...Fluorescence lifetime imaging microscopy method FLIM has been developed more recently and is the most rigorous...emergence of faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...Bioessays. 2012 May; 34(5):341-50. doi: 10.1002/bies.201100098. Sekar RB, Periasamy A. Fluorescence resonance...
  9. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging ... Overlapping  Overview: The overlapping strategy has a region of homology, typically ~400-1400 bp long...Drawbacks:  Generally an overlapping split vector has equal or higher transgene expression when compared...split AAV method While the hybrid dual vector system has a slight advantage over the overlapping and trans-splicing...efficiency of transgene expression from split vectors has been inconsistent. A greater understanding of basic...al., 2002 used an overlapping vector with 440 or 1000bp overlaps to deliver the alkaline phosphatase gene...
  10. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...kD TurboID, which has 15 mutations compared to BirA and the 28kD miniTurbo, which has the N-terminus domain...proteins in minutes rather than hours like BioID. APEX has allowed scientists to identify the proteomic map ...BioID was developed in 2012, proximity labelling has become a powerful technique for studying protein-...proximity-based labeling methods for interactome mapping. F1000Research, 8.  ...
  11. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...trimers, tetramers). Since then, multimerization has been observed to occur in many different kinds of...sequences. In some specific cases, multimerization has been observed to occur more frequently in plasmids... Ausubel, 1976). In fact, if your plasmid sample has a mix of different-sized multimers, you may even ...~3.6 kb and ~1.1 kb (lane 3). Uncut DNA (lane 2) has a band at <3 kb, which corresponds to the supercoiled...multimers is capillary gel electrophoresis (CGE), which has been used in plasmid vaccine and gene therapy development...Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/elps.201000061. PMID: 20589859. Levene SD. Analysis of DNA topoisomers...
  12. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ... cell mobility within these complex environments has never been easy and, for many years, researchers ...Intravital imaging powered by two-photon microscopy has uncovered host-pathogen interactions leading to the...proteins can be used in your experiments, experience has shown that some are more reliable and more efficient...important to finely tune the excitation wavelength as it has been shown that even a small incremental change of...this system called Brainbow, the Ubow mouse strain has been created to fate map Langerhans cells and follicular...with Toxoplasma gondii." PLoS Pathog 5.7 (2009): e1000505. PubMed PMID: 19578440. PubMed Central PMCID: ...
  13. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...invention in the late 1990s, Gateway cloning technology has become very popular as a rapid and highly efficient...attL-containing entry vector.   Pro Tip: Addgene also has ready-made entry clones available for many popular...Drawbacks. Clon Transgen 4:138. doi:10.4172/2168-9849.1000138 2. Hartley JL. Use of the Gateway System for...
  14. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...polyclonal cell population, meaning that the transgene has integrated in different locations in the various ...variety of dilutions and pick the population that has the most desirable level of expression. Over time...
  15. AAV ddPCR Titration

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...calculate the titer for each sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume...sealer and press the ‘Seal’ button. After the plate has been sealed, proceed to thermocycling. Thermal Cycling...
  16. CRISPR Guide

    Type
    Guide
    .... Cas9 nickase (Cas9n), a D10A mutant of SpCas9, has one active nuclease domain and one inactivated one... available. SaCas9 (from Staphylococcus aureus ) has a coding sequence ~1 kb shorter than SpCas9 while... SpCas9. FrCas9 (from Faecalibaculum rodentium ) has increased fidelity, and recognizes NNTA, a PAM sequence... create the adenine base editor. The TadA domain has also been further evolved to catalyze cytidine deamination...editing requires more optimization and typically has lower efficiency (< 50%) than base editing (> 90%...modifications to eliminate target gene function. It has also been used extensively to screen for novel genes... those for yeast or Drosophila . Although CRISPR has been less widely used in bacteria due to technical...
  17. Pipetting Protocol

    Type
    Protocol
    ...middle is hundreds, and the third is tens. Therefore, 1000 µL would read as 100 from top to bottom (as shown... flow rate into the pipette tip. Once the liquid has finished drawing into the pipette tip, lift the pipette...Anatomy of a Pipette This front view photo of a P1000 pipette shows the plunger at the top, which is used...tip off of the pipette. This side view photo of a P1000 pipette shows the volume adjustment ring right below...come in standard sizes: P2, P10, P20, P200, and P1000. The table shows the pipette with the appropriate...This image shows the P2, P10, P20, P100, P200, and P1000 as shown from left to right. Pipette Dispense Volume... 2 to 20 µL P100 10 to 100 µL P200 20 to 200 µL P1000 100 to 1,000 µL Each pipette should be used with...
  18. Colony Formation Titering Assay

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...mL) x dilution factor e.g., If the 1:100,000 well has 75 colonies, then there are 75 colonies /1.5 mL x...
Showing: 41 - 60 of 66 results