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We narrowed to 971 results for: lars

Showing: 281 - 300 of 971 results

  1. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in the femto-aptomolar range. For...fastest test that has been developed so far.  A similar method has been optimized to detect the SARS-CoV...molecule per milliliter can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C ...and will not give a positive result for highly similar viruses. Unlike Cas12a, Cas13a does not require...illuminator or UV light illuminator. Approach: Highly similar to the SHERLOCK diagnostic test, this test varies...identification using FnCas9 (FELUDA) Sensitivity: – 110 femtomolar  Time: Not mentioned, likely ~1-2 hours Equipment...Shravanti Suresh is a PhD candidate who investigates cellular aspects of CRISPR/Cas immune systems in the Sashital...

  2. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...Goedart, an assistant professor at the Section of Molecular Cytology and van Leeuwenhoek Centre for Advanced...Microscopy (University of Amsterdam). GFP is the most popular, most widely used genetically encoded fluorescent...fluorescent probe. Several factors contribute to the popularity of GFP including (i) fast and complete maturation...folded beta-barrel structure, followed by an intramolecular reaction that generates the chromophore (Tsien...Tsien, 1998). Importantly, this reaction requires molecular oxygen and as a consequence GFP remains non-fluorescent...Goedhart is an assistant professor at the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced...Kurt. "Genetically encoded fluorescent tags." Molecular biology of the cell 28.7 (2017): 848-857. PubMed...

  3. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...should be widely applicable to other targets. The modular system of linkers and spacers could enable many...single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with 10 minutes of glutaraldehyde crosslinking...previously-reported technique for precisely integrating large DNA sequences into a genome (Anzalone et al., 2022...deletion, replacement, integration and inversion of large DNA sequences with twin prime editing. Nature Biotechnology... (2024). Efficient site-specific integration of large genes in mammalian cells via continuously evolved...cells you are studying. Antibodies can help reveal cellular identities by targeting marker proteins that are...expressed in a given cell type. For example, glial fibrillary acidic protein (GFAP) is a glia-specific intermediate...

  4. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ... what your desired edit is. Large knock-ins Are you introducing a large insertion, something 20 base ... applicable to knock-ins both large and small is a software very similar to TIDE known as TIDER (Tracking...deletion that isn’t a multiple of three will do. A popular screening method for knockouts is Tracking of Indels...edit. Most dual guide systems generate a deletion large enough that it can be visualized by running your...guide system.   Fig. 2: Size screening for large deletion events.   Validating knock-in mutations... Pro tip: make sure your PCR amplicon isn’t too large so that you can easily visualize the size shift ...

  5. Sensing Neuronal Dopamine

    Type
    Blog Post
    ...dLight1, a molecular tool to detect and image DA in live animals (Patriarchi et al., 2018). Similar to calcium...the brain has been in vivo microdialysis of extracellular fluid (Darvesh et al., 2011). This technique... across the semipermeable membrane from the extracellular fluid into the perfused liquid. The probe then...sensor, dLight1.1 and dLight1.2, exhibit high (submicromolar) DA affinity and high-magnitude responses (increases...s lab has created the GRABDA sensor based on a similar strategy: a GFP variant bound to a DA receptor,...cyclase in caudate nucleus of rat brain, and its similarity to the “dopamine receptor”." Proceedings of the...

  6. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...and targeting power of CRISPR combined with the large sequence capacity of transposons open whole new ...’t cause double-stranded breaks and can deliver large sequences and even entire genes. Figure 1...template and transposon proteins (Tns) to integrate large sequences into genomic DNA downstream of gRNA-specified...Created with BioRender.com. CASTs quickly became popular bacterial genome engineering tools, with early ... al., 2019). Type I-F CASTs were found to be particularly useful. But they were barely effective at editing...., Gelsinger, D. R., Vo, P. L. H., Chen, A. T., Tolar, J., Osborn, M. J., Sternberg, S. H., & Liu, D. ...on Addgene.org Addgene's CRISPR Guide (Jump to Large Edits) CAST Plasmid Collection ...

  7. What the HEK?

    Type
    Blog Post
    ...293, the most commonly used line expresses SV40 large T antigen and is thus dubbed HEK-293T. Given that...older cell lines in the research community, many cellular research tools have been optimized specifically... viral production because they express the SV40 large T-antigen. This allows them to replicate viral plasmids...HEKs can also grow in suspension, which allows for large scalability, and can also be adapted for serum-free...two areas of research where HEK cells may be particularly useful due to their intrinsic properties. ...by mycoplasma and bacteria, but HEKs are also particularly vulnerable to viral infection. Yes, your cells...Additional resources on the Addgene blog Five Popular Model Organisms 10 Tips for Mammalian Cell Culture...

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

    Type
    Blog Post
    ...encoded tools, such as microbial opsins, to control cellular activities using light. In 2015, scientists combined...Charles Gersbach’s lab at Duke University developed similar systems based on the light-inducible heterodimerizing...effector (LACE) system, the Gersbach lab utilized a similar strategy to develop an optimized photoactivatable...system was able to recognize the same PAM and had similar targeting specificity as full-length Cas9 (flCas9...light on and off. As one might expect from the modular nature of Cas9, Nihongaki et al. showed that it... respectively. The use of far-red light is of particular interest since blue light has limited applications...that post-UV exposure it showed cleavage activity similar to the wild-type Cas9. This photocaging technique...

  9. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...this latest work, the team designed and screened a large library of iGluSnFR3 mutants, identifying the two...somatosensory cortex, while iGluSnFR4s produces larger signal-to-noise ratios in two-photon imaging, making... making it ideal for video recordings of large populations of synapses and deep-brain fiber photometry...most experiments. These new tools promise to be a popular choice for a variety of synaptic imaging applications...gene makes stable cell line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site...edits — single-base changes, small deletions, and large insertions up to 12 kb — with efficiencies between...isolating their associated protein interactome. Molecular Cell, 84(24), 4929-4944.e8. https://doi.org/10.1016...

  10. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...sometimes localize to unintended sites with sequence similarity to the on-target site. When a CRISPR-Cas system...frequency of 5%. Your experiment involves making a large sgRNA library to thousands of genes and targeting...targets by selecting a gRNA with low sequence similarity elsewhere in the genome. There are many predictive...Once you’ve selected a gRNA with low sequence similarity elsewhere in the genome to reduce off-targets...to consider your Cas enzyme. Even if there are similar sequences present in the genome, certain Cas enzymes...Candidate site sequencing Sites with high sequence similarity to the gRNA can be sequenced. These sites can...appropriate method for an activator or repressor Cas. Similarly, Cas ChIP-Seq would also identify all DNA binding...

  11. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    ...topics in biology, especially those at the molecular and cellular level. The reason for this is cost and ...it wasn’t until I started working in my first molecular biology lab in college that I really fell in love...limited budget. While there are more and more molecular biology-based activities making their way into... about operating the rest of the car as well. Similarly, cell free expression systems contain all the ...References 1. Huang, Ally, et al. "BioBits™ Explorer: A modular synthetic biology education kit." Science Advances... to learn simple lab techniques Check out our molecular biology reference pages Find plasmids from the...

  12. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ... by Alycia Bittner CRISPR pooled libraries are popular tools for genetic screens, enabling genome-wide...genome-wide interrogation of the genes required for cellular processes. For a successful screen, it is necessary...Schwiesow A leading cause of death worldwide, cardiovascular disease is driven by atherogenic lipoproteins...lipoproteins that accumulate within the vascular wall. ApoB, the main structural component of atherogenic lipoproteins...been shown to have a strong correlation with cardiovascular disease and indirect measurement of ApoB-LPs...to aid in the understanding and treatment of cardiovascular disease. Find the LipoGlo plasmids at Addgene...78% in body wall muscle cells. These changes were larger than previously reported for other organisms, and...

  13. Plasmids 101: How to Verify Your Plasmid Using a Restriction Digest Analysis

    Type
    Blog Post
    ...used to confirm the presence of an insert in a particular vector by excising the insert from the backbone...insert. You can search NCBI for YGOI to find the particular reference sequence if necessary. The example ...BamHI) are capable of cleaving sequences which are similar, but not identical, to their defined recognition... see 3 bands. This is due to the fact that the circular DNA takes on several conformations the most abundant... the DNA mass of comparably intense samples of similar size. Run the gel at 80-150V until you have good...the gel for longer to achieve good separation of larger DNA fragments. I hope these tips demonstrate ...More Plasmid Eductional Resources: Addgene's Molecular Biology Reference Find More Protocols and Videos...

  14. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...including the popular BL21(DE3) (Li et al., 2021). This pEcCas/pEcgRNA system works similar to the original...modifications, efficiency can be low, especially for larger modifications. What’s the solution to this inefficiency...recombinogenic than E. coli, so this system functions more similarly to CRISPR/Cas9 systems adapted for eukaryotes ...promoters are ubiquitous, they are not universal, particularly when it comes to non-model bacteria. The authors...engineer the bee gut microbiome. They created a modular system using a broad host range plasmid that can... specific locus. This approach provides a more granular level of control where individual bases can be...Bee Gut Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth...

  15. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    ...face very similar issues when forced to express in mesophilic host systems. The so-called “regular” temperatures...at suboptimal ones, which could allow even your regular E. coli to thrive in the bitter cold.  Since...vs. extremophiles Mesophilic organisms are your regular next-door microbes which grow optimally in moderate...psychrophilic enzymes for milk processing, fruit juice clarification, meat tendering, and oligosaccharide synthesis...the many proteins which are not compatible with regular mesophilic systems are the psychrophilic enzymes...incorporate cold-tolerance inducing genes within a regular mesophilic host system. This would make the host...

  16. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...reagents with the scientific community.   (click to enlarge) As we look ahead to the things to come, we wanted...total? Just over 49,000 times! Their long-lasting popularity illustrates the utility these tools have in many... The S. pyogenes Cas9 (SpCas9) has been widely popular in genome engineering. For use in human cell lines...plasmid. Indeed the first version is the 15th most popular plasmid of all time, but was found to have a point...publication but its name was subsequently changed to clarify that it does not contain the trademarked Tet-On...lentiGuide-Puro. pSpCas9n(BB)-2A-GFP (PX461) - Similar to pSpCas9(BB)-2A-Puro (PX458), pSpCas9n(BB)-2A-GFP...gRNA. It’s the former version of our sixth most popular plasmid pSpCas9(BB)-2A-Puro (PX459) V2.0. It contains...

  17. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    ...duties Staff scientist: similar role to a postdoc but more permanent Visiting scholar/student: visiting from...conferences. The Barry Goldwater scholarship is also a national scholarship that supports students interested...because they had close relatives suffering from similar diseases. Others were inspired by the imminent ...that you would be interested in can also help you clarify your goals and define next steps! More than serving...online for a while and observed. Apply for those scholarships that you don’t think you’re qualified for. Wayne...populations. Research universities often have scholarships and funds dedicated for undergraduates interested...

  18. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ...Common self-labeling tags Let’s take a look at some popular self-labeling tools: Tetracysteine: The first reported...variants run up to HaloTag11. HaloTag is now the most popular self-labeling tag in Addgene’s collection. CLIP-tag...on the protein of interest. Table 1: Summary of popular self-labeling tag systems Tag Size (amino... HaloTag for their “ticker tape” method for intracellular recordings, switching between different colored...Cornish, 2011). HaloTag and SNAP-tag are the most popular self-labeling enzymes in Addgene’s collection, ...that self-labeling protein tags are increasingly popular. Embrace the flexibility and switch things up for.... Two-colour live-cell nanoscale imaging of intracellular targets. Nature Communications, 7(1), Article...

  19. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...difficulties in amplifying a template, particularly from a large genome, you can recheck your primer design...plasmid cloning Read our molecular biology reference Find how-to videos for molecular biology and lab techniques...sequence, or if you want to amplify sequences from a large genome organisms like mouse or human. There are ...nanodrop to assemble the  fragments based on their molar ratio. Assembly of the fragments There are multiple...the backbone plasmid; a starting point could be a molar ratio of 1:2 (plasmid:insert), but this parameter...

  20. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced... can be excited many times, thereby producing a large number of emitted photons before they are destroyed...absorb the excitation light less efficiently. Similarly, decreasing the power of the excitation light ...these factors, environmental conditions such as cellular redox state and oxygen concentration may affect...Goedhart is an assistant professor at the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced...Marieke Mastop is a PhD candidate at the Section of Molecular Cytology (University of Amsterdam) where she develops...Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire Fluorescent Protein...
Showing: 281 - 300 of 971 results