We narrowed to 13 results for: TIM;
-
TypeGuide...conjugate or conjugation process. Sodium azide, an antimicrobial agent, can block some amines needed for conjugation...In those cases, thimerosal can be used as an antimicrobial agent. Other chemical interferences with conjugating...measurable light or color in response to a specific stimulus, can be conjugated to antibodies in order to allow...produces a colored product that accumulates over time which is relative to the amount of enzyme present...purchased already conjugated, which can save significant time in the lab. However, if the antibody you want is...ubiquitously expressed protein such as GAPDH or tubulin (sometimes referred to as housekeeping genes). This allows...protein or protein interactions in a specific timeframe or context. Immunoprecipitation (IP) IP uses immobilized...
-
Optogenetics Guide
TypeGuide...neurons upon exposure to light through direct stimulation of ion channels. Naturally occurring channelrhodopsins... nonspecific cation influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely ...been created via genetic point mutations, codon optimization, and chimeric fusion of domains from two different...engineered with enhancements such as: Human codon optimization Example: Halo Increased photocurrent amplitude...activate different neural populations at distinct times. Temporal considerations. Temporal precision is ...and promoter system used, there is an incubation time (days to weeks) until peak expression of the opsin...fused to Cry2 and the other half to CIB1, light-stimulated heterodimerization can reconstitute the protein... -
Plan Your Experiment
TypeGuide...be low, but generally higher than HDR pegRNA optimization can be challenging Prime Edit Repress or Interfere... and base editor variants often have different optimal editing windows. Be sure to refer to the literature...Using validated gRNAs can save your lab valuable time and resources when carrying out CRISPR experiments...gRNAs Once your target sequences are selected, it’s time to design your gRNA oligos and clone them into your...and express these components (Figure 2), and the optimal system will likely depend on your cell type. Figure...will work for your experiment) and the expression time (transient or stable). For plasmid and viral-based...best for your experiment. It may be necessary to optimize your delivery conditions, especially if you are... -
CRISPR Guide
TypeGuide... an optimized gRNA for your application (see: Plan Your Experiment ). In addition to an optimized gRNA... currently no defined measure of fidelity, so sometimes they will be referred to as increased fidelity...structured motif to enhance pegRNA stability PEmax - optimized PE architecture; increased synergy between PE ...compared to HDR. Prime editing requires more optimization and typically has lower efficiency (< 50%) than...integrase landing site ( attB ) and the other to stimulate DNA repair. This can result in more efficient .... gRNA design for CRISPR libraries is usually optimized to select for gRNAs with high on-target activity...activators and targeting promoter regions. You might sometimes see these referred to together as CRISPRi/a. dCas9... -
Molecular Biology Reference
TypeGuide...field of recombinant DNA technology have been optimized for their use of studying and manipulating genes...bacterial cell division is reduced (i.e., it takes more time to copy this extra DNA). Because of this reduced...form of another antibiotic resistance gene (this time, under the control of a non-bacterial promoter) ...origin of replication, and MCS. They are small and optimized to help in the initial cloning of a DNA fragment...developed the process termed Sanger sequencing, sometimes referred to as chain-termination sequencing or...Sanger sequencing requires a lot of DNA because the ultimate goal is to have a fluorescently tagged nucleotide...captures the fluorescent color that is emitted each time a base is added. Again, each base (A,C,T, or G) ... -
Cloning
TypeGuide... over the years to move around pieces of DNA. Oftentimes several approaches will work for any specific...specific DNA sequence, which can make it difficult at times to identify compatible restriction enzymes, that...behind a short scar in the DNA sequence and can be time consuming compared to other cloning methods. You...simplifying the process and reducing the amount of time required for cloning. To utilize this approach, ...proteins. Thus, you can clone your gene of interest one time into a donor plasmid (or acquire one that already...polymerase used, and the single A overhangs degrade over time, further reducing ligation efficiency. TOPO® ready... -
Adeno-associated virus (AAV) Guide
TypeGuide...viral replication (E4, E2a and VA). These are sometimes combined within the packaging plasmid. For a summary...occur, although at a low frequency which has been estimated to range from 0.01% to 3% depending on cell type... range of cell types in vivo. AAV-DJ has been optimized further for specific cell and tissue types, like... lab has developed plasmids containing SaCas9 optimized for their use in rAAV vectors . Read our blog ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear...protein (DBP) that enhances DNA replication by stimulating elongation and stabilizing the displaced single-stranded... -
Gamma-Retroviral Vector Guide
TypeGuide...start sites, such as promoters and enhancers. Many times, integration sites are found in proto-oncogenes.... of helper-free production cell lines also saves time in downstream purification, as extraneous plasmid...different retroviral vectors using a novel, CoDOn-Optimized gene for chimeric GALV envelope. Viruses , 13 ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear... -
Adenovirus Guide
TypeGuide...size of the DNA removed from the viral genome. Over time, subsequent generations of rAdV vectors have been...needing transgenes to express for a shorter period of time or when needing to package larger cargo. CRISPR-...vectors are also being used in cancer therapy to stimulate anti-tumor immune response in different ways, ... a given virus or infectious agent at a specific time point. Transduction The process of artificially ... -
Lentiviral Vector Guide
TypeGuide...assess viral integration (see Kim et al. (2021), Timms et al. (2018), and Wells et al. (2020) for examples..., A., & Pour, M. F. (2020). Concise review on optimized methods in production and transduction of lentiviral...https://doi.org/10.1261/rna.2192803 PMID: 12649500 Timms, R. T., Tchasovnikarova, I. A., & Lehner, P. J. ...virus post‐transcriptional regulatory element; stimulates the expression of transgenes via increased nuclear... -
Modular Cloning Guide
TypeGuide...conjunction with the MoClo Yeast Toolkit (YTK) to optimize secretion and cell surface display of proteins...are made with zCas9i, a high efficiency intron-optimized SpCas9-coding gene. MoClo CRISPR/Cas Toolkit for... -
Promoters
TypeGuide...RNA polymerase transcribes DNA to mRNA which is ultimately translated into a functional protein. Thus the... -
Chemogenetics Guide
TypeGuide...G-protein and activate neuronal firing through stimulating phospholipase C, which releases intracellular...