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We narrowed to 847 results for: PLE

Showing: 421 - 440 of 847 results

  1. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ...somewhat more reliable quantitation. Differences in sample treatment across methods can affect reported titer...that users keep titering method consistent across samples within a project. Targeting Your AAV Q4: Are there...for very high MOI on top of very high coverage. Please see this paper for more information about PHP.eB... will very likely cross-react with each other. Multiple labs have tried to transfer PHP.eB's enhanced ...which may be linked to epigenetic changes, such as multiple sclerosis?  A: Basic science and preclinical researchers...

  2. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    Published
    Feb. 8, 2022, 2:15 p.m.
    ... For example, setting up your pipette tips to match your plate layout can help you track samples as you...change variables to fix a failed experiment.  For example, I used to constantly underestimate time needed... and make a checkmark next to each step as you complete it. Yes, this can be incredibly tedious, but it...remember what's been added and what hasn't. Developing simple habits like these can consistently reduce errors...

  3. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    Published
    May 25, 2017, 2:30 p.m.
    .... This luminescent technology is based on the principle of bioluminescence resonance energy transfer (...photobleaching/phototoxicity, they successfully imaged multiple subcellular compartments over several minutes ...improve long-term imaging, which is limited by depletion of luminescent substrate, they synthesized diacetyl... handy. This approach is especially suited to complement optogenetic tools, where light used for excitation...external light source. Therefore, especially thick samples may not be appropriate for use with Nano-lantern...

  4. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    Published
    Jan. 25, 2024, 2:15 p.m.
    ...Cis/trans activity, PAM requirement, gRNA, and multiplex ability Experimental guidelines and applications... the cis and trans activity of each entry. For example, SpyCas9a (one of the most frequently used Cas ...you need more nitty-gritty details — how can I multiplex this enzyme? What other RNA components are required.... For each enzyme, it will also show you how multiplexed arrays would be designed, if that’s your end ...details to refine your search (trans activity, multiplex ability, gRNA design, etc.). With this feature...

  5. Viral Vectors 101: Biosensors

    Type
    Blog Post
    Published
    July 6, 2023, 1 p.m.
    ...present in the system at a given point in time. For example, if ATP was the molecule of interest (analyte),...depending on the biology you are studying. For example, calcium dynamics in the cytosol and sarcoplasmic... have been tested in vivo, in tissue preps, in multiple species, and in cell culture models. Some of them... them have even been expanded for multiplexed imaging for advanced monitoring of different factors, in...all have specific sensors designed for them. For example, this class includes tension monitors associated...

  6. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...that use reverse transcriptase to synthesize complementary DNA copies of their genome upon infecting a ... These viruses have a cargo capacity of ~ 8 kb, ample room for most experimental needs, and can transduce...administration, such as in mouse experiments. AAV also has multiple serotypes (12 naturally occurring and more synthetic...enough for many applications, but in some cases multiple AAVs (co-infection), an alternative virus, or ...applications these viruses are good for, we have plenty of information on that too. May your titer always...

  7. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    Published
    March 21, 2023, 1:15 p.m.
    ...applications, while electroporation of a human, for example, isn’t a reasonable option. Since viruses used...used in labs are derived from viruses that infect people in nature, extra safety measures, like replication...therapies have already been approved in the US for multiple diseases, including hemophilia B and spinal muscular... immunodeficiency virus type 1 preintegration complexes.” Proc. Natl. Acad. Sci. U.S.A. 89 (1992) 6580... Multiple Authors...

  8. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ... Methylases involved with these processes (for example Dam and Dcm methylases) are independent from the...conversely, activating enzymes such as DpnI. For example, cleavage by XbaI may be blocked due to methylation...those sites will be methylated. Using the same example plasmid as above (which has the XbaI site blocked...methylation by altering your choice of bacteria. For example, if you must use a restriction site that will be...produce DNA that is unmethylated at those sites. Please keep in mind that dam-/dcm- strains may have an...

  9. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    Published
    June 2, 2016, 2:30 p.m.
    ...labels, since I often work on multiple projects in a given day. For example, instead of naming a file “output...experiments can be found in each folder. This can be a simple .txt document. Record the name, date, and location...folder: You will often use the same data files for multiple analyses. Rather than copying these files to your...result was, so try not to do this. These are the simple rules that I try to follow in my own research. ...

  10. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    Published
    April 10, 2018, 12:56 p.m.
    ...of the sample due to the smaller size of RANbodies compared to antibodies. Staining with multiple HRP-RANbodies...used at a time. Samples can be sequentially stained with two HRP RANbodies if the sample is stripped. HRP-RANbodies...process that’s difficult for individual labs to complete. The nanobody based RANbody platform from the ...

  11. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...Assembly, Golden Gate etc.), and try to find the simplest way to do it (i.e. avoid assembling too many fragments...methylation like XbaI, ClaI, etc. If you are doing multiple digestion be sure that the buffers and temperature...Remember to quantify the DNA concentration of all the samples by gel or by nanodrop to assemble the  fragments...molar ratio. Assembly of the fragments There are multiple ways you can assemble the different parts of a...plasmid. Many vectors contain sequences that are complementary to standard sequencing primers. If not, design...

  12. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    Published
    Nov. 30, 2021, 2:15 p.m.
    ...ill-defined. In this article, I try to clarify what people mean when they ask about “problem solving skills...problem.  Step Three: Decide on a strategy and implement it.  Step Four: Assess how effective your strategy...the cause. To test the most likely cause, you implement an experiment designed to fix the problem. Then...modifications, and ultimately decided on the changes to implement. As with any skill, there will always be room ...Know your strengths! Research experience presents ample opportunities to practice problem solving skills...

  13. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...outlined in Table 1.   Of course, this is for the simple insertion of a PCR product into a vector. The time...calculate, in both our 2018 and 2023 poll, most people said it took them more than 22 hours, so at least...published in. That’s also a great way to get to know people doing interesting and relevant work in your field...That way, you can run one PCR, one gel, etc. for multiple plasmids at once. Over time, this can save you... Multiple Authors...

  14. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...is guided by an RNA molecule, the gRNA, to a complementary site in the genome where it will proceed to ...degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction. Cas3 is ...that Cas14 also indiscriminately cleaves non-complementary ssDNA when engaged by its target sequence. Why...used in lieu of siRNA or shRNA for efficient, multiplexable RNA knockdown. Cas13 has also stirred up interest... the other Cas family members make for useful implements in the scientist toolbox. If your genome engineering...

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

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...ccdB aspect, Gateway takes advantage of the same principle that cells will not propagate while expressing...system contains ccdB. A successful insertion will completely replace ccdB with the investigator’s insert of...interest.   CcdB resistant E. coli strains complete the system But ccdB alone isn’t the only key to...most appropriate method for your own experiments. Please check out Addgene’s Choosing a Molecular Cloning...protein involves poisoning of DNA-topoisomerase II complexes.” J Mol Biol. 1992 Aug 5;226(3):735-45. PubMed...

  16. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    Published
    July 21, 2016, 2:30 p.m.
    ...conduct research that fails less and helps more people, we need to take sex into account. Gendered Innovations...s Health Initiative Hormone Therapy Trial, for example, found that the study saved lives. Reduced use ...analysis and 2) provides case studies with concrete examples of how gender analysis leads to discovery and ...experiments—which can lead to failed research. For example, an international research team from Norway and...studies that draw from a body of research that is incomplete (because sex was not analyzed) may harm humans...

  17. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...ever like to write about a recent plasmid deposit please sign up here.  Here's what you'll find in this ...expanding our ability to study and understand the complex interworking of the 86 billion neurons in the human...Using genetically encoded fluorescent proteins coupled with Fibronectin intrabodies generated with mRNA...speed across these different cell types. In one example, H2B-FT was expressed in murine B cells (BaF3),... the plasmids at Addgene The Cas Hybrid for Multiplexed Editing and screening Applications (CHyMErA) ...

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

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ... the xanthine produced via complete demethylation of caffeine. For example, the Barrick lab was able to...ever like to write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode...living biosensor for measuring caffeine content in samples. The Barrick lab and the UT Austin undergraduate... to grow this strain in minimal media supplemented with various caffeinated beverages, but not with caffeine-free...

  19. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ... the mutation or replacing the gene. A notable example is the treatment of Severe Combined Immune Deficiency...normally functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside...based delivery techniques may hold the key. In the simplest form, translating gene therapies from conceptual...already have ways to prevent AAV infection. For example, prevalence of neutralizing antibodies against ...: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates gene targeting and enhances endonuclease...

  20. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...CRISPR is a simple and versatile tool for genome engineering, but its utility is dependent on its ability...tiny DNA cutting ninjas that deliver CRISPR to multiple types of targets: primary cells, embryos, and ...helps form DNA and retroviral-particle or VLPs complexes. The team found that "all-in-one" Nanoblades facilitated...their CRISPR cargo into the zygote. As a proof-of-principle, the team used Nanoblades to disrupt the tyrosinase...no detectable editing. Additionally, off-target spleen editing was not detected and Nanoblade injection...
Showing: 421 - 440 of 847 results