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We narrowed to 490 results for: ARA-2

Showing: 361 - 380 of 490 results

  1. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    ...responsible for that differentiation.   Fig. 2: Single cell RNA-seq clustering of cells transduced...road map, and what was known about previously characterized TFs, the Zhang group were able to classify unknown...studied TFs, this functional association of the uncharacterized TFs is a useful finding. Transcriptional program...

  2. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...and open a new conversation about GMOs.  Challenge #2: Technical obstacles to plant synthetic biology In...transformation to testing and a lack of well-characterized genetic tools make it challenging to engineer...understood. Additional work has also identified and characterized mechanisms and components of the circadian rhythm...

  4. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...divided by the off-target activity. Figure 2: Specificity is the measure of off-target activity...experimental approaches. One example is CasX, characterized by the Oakes and Doudna Labs, which is both ...research and potentially treatments. Cas12b, characterized by the Zhang (Strecker et al., 2019) and Li ... relative to Cas9. As scientists continue to characterize CRISPR nucleases from a variety of sources, ...

  5. Antibodies 101: Isotypes

    Type
    Blog Post
    ...IgM, IgD, and IgG are deleted.     Figure 2: Isotype switching requires DNA recombination of the...isotype? An isotype determines several key characteristics of an antibody as well as the role it plays... regions result in several unique physical characteristics of these five isotypes, such as the number ...IgG4. IgE IgE antibodies help protect against parasites but also play a role in allergies. They are found...

  6. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication ...from Pseudomonas aeruginosa to cleave 4 gRNAs separated by 28 base Csy4 recognition sites from a polycistronic...protoplasts. Each pair of gRNAs targeted genomic sites separated by 350-750 bp, allowing them to examine the frequency...different biological systems. Although Xie et. al characterized the system in plants, PTGs will likely work ...

  7. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...extraction solution, runs the protocol, then uses around 2–4 µl in each 50 µl PCR reaction. The gDNA extracted... early onset Parkinson Point mutations. Cell, 146(2), 318–331. https://doi.org/10.1016/j.cell.2011.06.019...system, developed by the Zhang lab, which has comparable efficiency to the optimized chimeric design but...for PCR and surveyor. Q15: I used DNA polymerase Takara Ex Taq™ DNA Polymerase for my genomic PCR, but ...successfully used Herc II or Kapa Hifi in PCR. Maybe this Takara enzyme is not very robust in this case for EMX1...processing of the crRNA) is also expressed from a separate promoter. In the pre-processing form, the crRNA...

  8. Antibodies 101: Epitope Tags

    Type
    Blog Post
    ...is the most charged, with 5 negatively charged and 2 positively charged amino acids. FLAG tag has been ...the need for an antibody. Tags can enable the separation of detection of a mutant protein vs. endogenous...

  10. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

    Type
    Blog Post
    ...still depolarize neurons at lower MOIs. Figure 2: ChRgers are delivered by intravenous injection into... “designer” channelrhodopsins with desired characteristics. The model was trained to predict if a channelrhodopsin...these remaining sequences were selected for characterization with patch clamp electrophysiology. All thirty...named ChRger1, ChRger2, and ChRger3, were then characterized further by expression in cultured neurons and...performed well in a tissue culture dish, that doesn’t guarantee their success in vivo. Therefore, the Arnold and...

  11. RNA Extraction Without A Kit

    Type
    Blog Post
    ...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method...gradients and ultracentrifuged for up to 20 hours to separate the intact RNA from DNA and proteins. Although...and Sacchi showed that RNA could be effectively separated from DNA and proteins by a simple extraction protocol...thiocyanate solution. However, instead of RNA separation using cesium-chloride gradients, water-saturated...ultracentrifugation!), the layers of phenol and chloroform separate, and RNA is retained in the top, aqueous layer...

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

    Type
    Blog Post
    ...main decisions to make: 1. Which modification? and 2. Which viral vector should be used? Modifying gene...need to be given some time to adapt to a testing apparatus. General stress levels of the test animal may ...the humans’ audible spectrum. The experimental apparatus should be cleaned thoroughly between animals as...beginning. Variability source Standardization parameters Animal Strain, age, sex, housing conditions...order to maximize possible effects. Injection Parameters: The injection needle should be as thin as possible...

  13. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ..., but at a lower efficiency (~20%). Figure 2: Schematic of multicomponent assembly with SLIC. Multiple... other dNTPs. Once the vector and insert are (separately) digested to create 10–12 base “chewed back” ... T4 polymerase (left branch); by annealing two separate PCR products (middle branch); or by incomplete...

  14. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ... and transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above...for processing several sequencing reactions in parallel. We have a few tricks to simplify the process....saves us the step of purifying the DNA and gives comparable success rates and sequence quality, which is ...

  16. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...10.3389/fmicb.2020.607028  Riley, R., et al., Comparative genomics of biotechnologically important yeasts...

  17. Components of CRISPR/Cas9

    Type
    Blog Post
    ...Without binding to the guide RNA, Cas9 cannot cut. 2. Bind to target DNA in the presence of a guide RNA...bacterial genomes consist of repeated elements separated by unique sequences. When researchers first discovered...

  18. How Addgene Creates Content

    Type
    Blog Post
    ...and in line with Addgene style.        Fig. 2: Reference image (left) provided to our graphic designer...the antibody guide always gets an expert review, separate from the author, and a second review from someone...

  19. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...to visualize multiprotein complexes.  Fig. 2. Multiplex immunolabeling with subclass-switched recombinant...and validation pipeline. (B) Schematic shows the separate elements of the R-mAb expression plasmid involved...

  20. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...two options to begin your plasmid deposit. Figure 2: Choose to submit plasmid(s) online or by using a ...format. Figure 6: Add sequence, maps, and files. Step 2: Gene and Insert *Note: This page will not appear ... into it. You will have the option to indicate separate promoters, tags, or cloning information for each...
Showing: 361 - 380 of 490 results