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We narrowed to 576 results for: BASE

Showing: 121 - 140 of 576 results

  1. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this syringe-based method...Compared to a commercial magnetic bead-based kit, this cellulose-based method was faster, cheaper, and had... Addgene follows a similar workflow as a column-based plasmid extraction kit. First, you lyse the bacteria...

  2. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...are many protein-based toxin-antitoxin systems. One of the most well known protein-based toxin-antitoxin...counter this, methyltransferases add methyl groups to bases at specific nucleotide sequences, preventing the...system that is regulated by antisense RNA, which base pairs to cognate mRNA to inhibit its translation...Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition." Molecular cell 11.4...

  3. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ... let the virions out. We recommend using a Tris-based lysis buffer and sonication.  The obtained virions...particles that can transduce cells, which can vary based on target cells and can be altered by freeze/thaw...experimental outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers, meaning...post.  We generally do not recommend using ELISA-based titering assays, since those will quantify both ...Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based Disinfectants. Human Gene Therapy, 32(13–14), Article...

  4. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...for studying colloidal gold (1).  The method was based on using a thin plane (sheet) of light generated...microscopes, optical sectioning of a specimen is based on discriminating the out-of-focus reflected light... combining Ultramicroscopy with tetrahydrofuran-based tissue clearing (13,14). A few years later, Karl...Deisseroth’s lab developed CLARITY, which is a hydrogel-based method that also allows antibody penetration for...recently developed AAV (adeno-associated virus)-based fluorescent labeling methods that can be used with...

  5. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to analyze gut microbiota...microbiome, James Collins’s lab developed a paper-based method that can be used to detect microbes and biomarkers...As mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect mRNA transcripts...Takahashi, Melissa K., et al. "A low-cost paper-based synthetic biology platform for analyzing gut microbiota...

  6. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail from the second tRNA... cassettes (considering the gRNA itself is ~100 bases) limit the number of gRNAs that can be packaged ...Pseudomonas aeruginosa to cleave 4 gRNAs separated by 28 base Csy4 recognition sites from a polycistronic transcript...

  7. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...LoxP (locus of X(cross)-over in P1) sites are 34-base-pair long recognition sequences consisting of two...event. This event can have three general outcomes based on the location and orientation of the loxP sites...above, a genetic modification can be restricted based on experimental constraints. This has been used ...asymmetric spacer "NNNTANNN", where "N" indicates which bases may vary from the canonical sequence. Among these...

  8. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...options come in two categories: protein-based and chemical-based. Before selecting one, check your antibody...concentration. That makes the choice easy! Protein-based blockers include dry non-fat milk or serums like... of blocker, you can also use gelatin.  Protein-based blockers are inexpensive and easily made in the ... tip! If you get a speckled image, your protein-based blocker is either degraded or not completely mixed...mixed together.   Commercially available chemical-based blockers, while much more expensive, are consistent...to another. Since antibody binding varies wildly based on proteins, and their differing amounts of available...Smaller dynamic range; no multiplexing Protein-based blocker Degrades quickly Wet transfer Increased...

  9. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    Published
    Oct. 11, 2016, 2:30 p.m.
    ...years, deaminase-mediated base editing (cytosine base editor or adenine base editor) and reverse transcriptase-mediated...in Zhu et al., 2019). Both cytosine base editors and adenine base editors have been developed for plants...Belhaj et al., 2013). Nuclease dead Cas9 (dCas9) based activators (such as dCas9-VP64) or repressors (dCas9...

  10. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    Published
    July 10, 2018, 1:08 p.m.
    ...scientific field-based solutions or open source solutions. In comparison to paper-based record keeping,...notebooks and researcher's needs The familiar paper-based lab notebook is still a standard approach to record-keeping... or brainstorm, but we have to admit that paper-based record keeping barely copes with the growing amounts...future? As most payable software solutions have user-based pricing, it is good to know how many of your lab...

  11. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...day, you should be happy to learn that many DNA-based reagents are named for soup ingredients! From pSOUP... intracellular RNA dynamics. Aptamers are 10-100 base nucleic acid oligos that bind with high affinity...that control the expression of coding RNA. Spinach-based riboswitches can be used as stable metabolite sensors...mimic of green fluorescent protein by fluorescence-based selection and directed evolution." Journal of the...PMC4244833. Kellenberger, Colleen A., et al. "RNA-based fluorescent biosensors for live cell imaging of ...

  12. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    Published
    Jan. 26, 2023, 2:15 p.m.
    ... modifications (~1-50 base pairs). Small edits require as little as 30-50 bases for each homology arm,...arm, but keep in mind these numbers may vary based on your locus of interest and experimental system.  Synthesizing...  Synthesizing ssODNs longer than 200 bases can be difficult, thus dsDNA plasmids are generally preferred...Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to Improve HDR Efficiency ...

  13. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...process. Each variant is assigned a functional score based on its library frequency before selection compared...libraries of DNA fragments that contain variable bases (NNN) at any given codon site of interest can be... technique described in Wrenbeck et al is a PCR-based approach for generating a customizable comprehensive...primer set contains three consecutive randomized bases (NNN) at a given codon site and multiple primer ...Keith E J Tyo, and Timothy A. Whitehead. "Plasmid-based one-pot saturation mutagenesis." Nature Methods ...contain random synthesis errors, especially single basepair deletions. Therefore, Gibson assembly is better...codons that can be mutated? 88 codons, or 264 basepairs, was the maximum length tested by Wrenbeck et ...

  14. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...latency (Bialek et al., 2016). Additionally, CRISPR-based activation can be made inducible upon environmental...through all the troubleshooting required for Cas9-based activation. CRISPRa for genome-wide studies One ...larger scale of activation allows for exploratory-based research as well. For example, CRISPRa can be used... interactions. Additionally, large scale CRISPR-based screens can be used to gain insight into protein...date his group has develop several popular CRISPR-based tools for altering DNA sequence or regulating gene...

  16. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...will focus on one of the most common, purification based on the polyhistidine tag. What is the polyhistidine...tag was developed as a means to purify a protein based on the affinity of the imidazole ring of histidine...Bornhorst 2000).  One downside of polyhistidine tag-based purification, however, is the propensity for nonspecific... tags can also be used as a target for antibody based assays. Antibodies are useful tools for studying...

  17. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ...amino acid linker to use. These decisions were made based on known functional aspects of the protein gleaned...sequence into which it is being inserted: one thousand base pairs on each side, in fact. Later, when we transfect...precautionary measure, it’s also good to make a few single base pair mutations in crRNA target sites found within...endogenous gene tagging systems. Addgene’s CRISPR-based Protein Tagging collection includes systems for ...

  18. Fluorescent Proteins 101: Fluorescent Biosensors

    Type
    Blog Post
    Published
    May 18, 2017, 2:30 p.m.
    ...Reporter signals may be electrochemical or light-based, with luminescent and fluorescent reporters being... domain induces changes in a fluorescent signal based on ligand presence or changing cellular conditions...Method Notes Quantification of a process Plate-based assays (plate reader) Detect the average amount...

  19. Twenty Years of Sharing Science!

    Type
    Blog Post
    Published
    Jan. 8, 2024, 3:25 p.m.
    ...repository for DNA-based materials, including our popular kits and libraries. This DNA-based materials repository...

  20. CRISPR in the Clinic

    Type
    Blog Post
    Published
    Nov. 2, 2023, 1:15 p.m.
    ...initiating a base editor-based clinical trial for SCD in 2022 and Editas Medicine starting a Cas12-based clinical... today. Below we will review some of the CRISPR-based editing therapies that are currently approved or...
Showing: 121 - 140 of 576 results