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We narrowed to 492 results for: ara-2

Showing: 161 - 180 of 492 results

  1. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...calcium biosensor, so that the (1) binding of calcium, (2) allosteric rearrangements of the sensor, and (3) ...user-controlled illumination with violet light and (2) exposure to increased intracellular calcium. It has...dynamics in vivo. Neuroscience Research. 2021 Aug 1;169:2-8. Suzuki J, Kanemaru K, Iino M. Genetically encoded...experiment with a sensor that is tuned to your parameters.   Kinetics How fast is the process you would...used calcium reporters with different kinetic parameters. There are three GCaMP types based on their kinetics...available with improved kinetic and sensitivity parameters. Sensitivity and Localization The calcium concentrations...to go. Pro tip: Vectors with ready-to-use AAV preparations are labeled with the green virus particle on...

  2. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    ...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit...safer viral vectors that can be used in BSL-1 or BSL-2 labs. Viral vectors are viruses that have been genetically...genes for the required proteins are packaged in separate plasmids and expressed via cells cultured with...

  3. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12... before beginning your experiments. Figure 2: Strategy for assembling barcoded combinatorial gRNA...PMID: 25114216. PubMed Central PMCID: PMC4151723. 2. Doench, John G., et al. "Optimized sgRNA design to...has been successfully applied to functionally characterize combinatorial gene knockouts generated via multiplexed...antibiotic-resistant bacteria enabled by massively parallel combinatorial genetics." Proceedings of the National...PMC3972032. 10. Wong, Alan SL, et al. "Massively parallel high-order combinatorial genetics in human cells...

  4. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ... was first discovered and characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching...neoblast cells and the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver...Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado, The American Biology...riboprobes are created, the WISH protocol involves preparation of the tissue sample, several washes, and temperature...PMC304099. Additional Resources on the Addgene Blog Arabidopsis in Education Learning Synthetic Biology with ...

  5. OCaMP: A New Calcium Indicator for Neural Imaging

    Type
    Blog Post
    ...the fluorescent protein mOrange2 (Figure 2). Figure 2: OCaMP structure and mechanism of action...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...Whether you're working with cell cultures, slice preparations, or in vivo imaging, there's a version that ...the science behind OCaMP, including detailed characterization as protein or in neuronal cultures, zebrafish...

  6. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...point could be a molar ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to... working with is correct. It's also best to use 1-2 ug of the vector for digestion. PCR Design primers...colonies (5-10) and grow them in a small culture volume (2 or 3 ml) containing the corresponding antibiotic, ...to be strategy specific, there are few general parameters that you can adjust: temperature and time of ...

  7. The time and cost required to make a plasmid

    Type
    Blog Post
    ...required to make a plasmid. In 2023 (and at closer to 2 million plasmids shared than not!), we think it’s ...time does it take to make a plasmid? Figure 2: Survey results from a Twitter poll run in 2023, n...go wrong, replace them for free. Plasmids ship out 2-3 days after ordering, making it a toss-up as to whether...relevant work in your field! However, there’s no guarantee you’ll get a response on the first try (or any...at a time, try constructing several plasmids in parallel. That way, you can run one PCR, one gel, etc. ...

  8. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...or non-homologous end joining (see Figure 2).  Figure 2: Schematic representation of integration...due to insertional mutagenesis (Howe et al., 2008). 2. Transient expression of transgenes/shRNAs If you ...vectors: progress and applications. Gene Therapy, 17(2), 150–157. https://doi.org/10.1038/gt.2009.135. PubMed...efficiently during infection. Journal of Virology, 70(2), 721–728. https://doi.org/10.1128/JVI.70.2.721-728.1996...our knowledge here, the Pol gene encodes three separate enzymes that are proteolytically processed from...

  9. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ... used. Follow the manufacturer’s instructions.   2. Isolate Your Insert and Vector by Gel Purification...DNA concentration alone. One method is to conduct 2 ligations for each plasmid you are trying to create...containing the ligation mixture with backbone alone; 2. Transformation containing ligation mixture with insert...colonies than the vector + insert plate (unsuccessful 2), or roughly equivalent numbers of colonies on each... or equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid...slower than smaller linearized fragments. You can separate your backbone away from any inserts cut out of...

  10. CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping

    Type
    Blog Post
    ...of other Cas proteins found in the so-called class 2 CRISPR-Cas systems. There are 24 variants of the cas14...the presence of infectious organisms (ex: SARS-CoV-2) and genetic mutations. By providing a gRNA specific...Department of Energy’s Joint Genome Institute for uncharacterized genes that were proximal to a CRISPR array ...presence of a predicted RuvC nuclease domain, characteristic of CRISPR-Cas enzymes. In contrast to other...

  11. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...research experiences to undergraduate students (1, 2), but educators face significant challenges in designing... by western blotting in other experiments (Figure 2).   Complementation reveals conservation of function...Biology Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology...Functional characterization of the Saccharomyces cerevisiae genome by gene deletion and parallel analysis...cerevisiae (Figure 1) occurs through a well-characterized pathway with multiple enzymatic steps (5). In.... cerevisiae met deficiencies (manuscript in preparation), indicating that Met protein functions have ...

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

    Type
    Blog Post
    ...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates...common cytokine receptor gamma chain (c) and is characterized by a lack of immune cell development and function...first discovered as a contaminant of adenovirus preparations. It is an icosahedral, non-enveloped virus that...repair via homology directed repair (HDR). This characteristic makes AAV vectors especially amenable to correcting...caused by mutations in the gene RPE65. It is characterized by childhood onset of blindness and was thought...

  13. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    ...possible when each of the XFP genes is introduced at ~2 copies per cell per color, following a Poisson distribution...(prepared with Addgene #104110 - #104112) (Figure 2). We’ve also created constructs with containing chemical...AAV titer - The XFP genes have to be introduced at ~2 copies per cell per color to achieve the highest color... levels of XFPs - Reduce the amount of tTA. Paradoxically, too much tTA leads to a reduced expression ...

  14. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    ...human transcription factors such as PAX7, PAX6, AP-2 alpha and ISLET-1. In addition, we distribute monoclonal... example through the use of less toxic adjuvants; 2) generating complex antibody chips that allow assessment...variable, allowing exquisite specificity. This characteristic allows monoclonal antibodies to be used as ...

  15. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly analyzed AAV-68544, we were...look for potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual...Léger A (2015) Advanced Characterization of DNA Molecules in rAAV Vector Preparations by Single-stranded Virus...especially important when producing multiple vectors in parallel because the risk of accidental errors increases... be used to identify DNA contaminants in AAV preparations and provide information on these DNA species...pioneering the systematic use of this new QC assay to guarantee that you receive the best AAV vectors for your...Identity of Recombinant Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy ...

  16. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227... Find plasmids from this publication at Addgene. 2. Kleinstiver, Benjamin P., et al. "Genome-wide specificities...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674...also called Cas12a. In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-...Until recently, two major elements of Cpf1 characterization were missing: on-target and off-target editing...efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency of two Cpf1 orthologs...lower than that of the widely used SpCas9 and comparable to SaCas9. As seen with Cas9 orthologs, Cpf1 ...

  17. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ... fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly to...PMID: 24194740. PubMed Central PMCID: PMC3810607. 2. Bajar, Bryce T., et al. "A Guide to Fluorescent Protein...PubMed PMID: 10500161. PubMed Central PMCID: PMC18018. 2. Miyawaki, Atsushi, et al. "Fluorescent indicators...various papers to make the decision easier. 1. Müller, Sara M., et al. "Quantification of Förster resonance ...donor and acceptor fluorophores are fused to two separate proteins. (In contrast, intramolecular FRET probes...error in suboptimal or undetectable FRET, well-characterized FRET reference standards can be used to validate...

  18. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...procedure to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As...Primer design Using specific primer design (Figure 2), IVA cloning can be used to perform any plasmid modification... for each type of modification is shown in Figure 2, and is discussed in more detail below. The main requirement...Biotechniques 10.1 (1991): 62-66. PubMed PMID: 2003926. 2. Bubeck, Peter, Monica Winkler, and Wilfried Bautsch...another simply amplify the vector and insert with separate primer pairs in the same PCR. Add homologous sequences...

  19. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...therapy had fallen - plummeted really -  from grace 2 years prior with the death of Jesse Gelsinger in an... the whole field to a grinding stop and only now, 2 decades later, is it finally back in the limelight...the other hand, have broad tropism in the eye. AAV-2, the first identified serotype and the one used in...reading 1. FDA approves gene therapy for blindness 2. Gene Therapy emerges from disgrace to be the next...occurring animal model, the RPE65−/− dog, was characterized and found to suffer from early and severe visual...

  20. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important ...1996 Apr 12;272(5259):263-7. PubMed PMID: 8602510. 2. A third-generation lentivirus vector with a conditional...CH, Humeau LM, Rebello T. J Gene Med. 2013 Feb;15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment... Banasik MB, McCray PB Jr. Gene Ther. 2010 Feb;17(2):150-7. PubMed PMID: 19847206. Additional Resources...Mulder GJ, Polack A, Bank A, Soulier J, Larghero J, Kabbara N, Dalle B, Gourmel B, Socie G, Chrétien S, Cartier...
Showing: 161 - 180 of 492 results