We narrowed to 986 results for: IGH@
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TypeProtocol...reagent you will be weighing out, a weighing boat or weighing paper to contain the reagent while on the...that you are weighing by looking for a weight range on the scale. Make sure that the weight of the material...out the material you are weighing onto the weighing boat or paper. If you weighed out too much material,... up. Discard the weighing boat or weighing paper in the trash if the material weighed is not biohazardous... Protocols Weighing Reagents Weighing Reagents Protocol Intro ... to handling the materials. Equipment Weighing boats or weighing paper Balance Scoopula or spatula Container...button. Before weighing out your reagents, determine the amount that you need to weigh. Gather the reagent...
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Using a Light Microscope Protocol
TypeProtocol...The route the light follows from the source to your eyes is called the light path . Light travels from ... Protocols Using a Light Microscope Using a Light Microscope Intro ... to encounter: the compound light microscope. As the name suggests, light microscopes take advantage of...compound light microscope with the main components labeled: Figure 1: Diagram of a compound light microscope...from the light source through a condenser that helps concentrate the light onto the sample, and then through...objective! Once on the higher objective, you will likely need to re-adjust lighting and focus. Increase ...basics of how to use a light microscope. Protocols... -
SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas
TypeBlog Post...coronavirus using CRISPR-based SHERLOCK (Specific High sensitivity Enzymatic Reporter unLOCKing)... under either no excitation light, UV light or LED blue light. Highlights: This is a one-pot, one-temperature...heat block, LED blue light illuminator or UV light illuminator. Approach: Highly similar to the SHERLOCK...captured by an antibody further along on the strip. Highlights: Of the 83 known COVID-positive samples tested... extraction but that may work only if there is a high enough (105 copies per microliter) viral load in...lateral-flow readout slip that allows rapid detection. Highlights: Cas13a is not catalytically active in the presence... duplex and will not give a positive result for highly similar viruses. Unlike Cas12a, Cas13a does not... -
Optogenetics Guide
TypeGuide...Microbial Opsins Opsins are light-gated ion channels or pumps that absorb light at specific wavelengths. ...respond to different wavelengths of light, like blue or yellow light. These various opsins also initiate...also a blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2 has higher conductance...Channelrhodopsin from Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum... 566 ArchT Light-activated outward proton pump from Halorubrum strain TP009. Improved light sensitivity...binding domain, allowing light to activate transcription. Phytochromes and light oxygen voltage (LOV) domains...) BLUF domains bPAC Light-activated adenylyl cyclase from Beggiatoa sp. for light-induced cAMP modulation... -
Antibody Guide
TypeGuide...the Fab fragments contain highly variable regions, shown in light blue and light orange, that bind to a ...are light-sensitive molecules which emit a specific wavelength of light after excitation by light of a...pair the brightness of the fluorophore with the abundance of the antibody target. If a bright fluorophore...molecular tools used in the lab to specifically and tightly bind to other proteins. Antibodies, produced as...-efficient way to produce antibodies, with the highest variability in antibodies produced. Cells - Antibodies...protein chains: two large heavy chains and two small light chains. These four domains come together to form... to be produced in a single individual, giving a high probability that an antibody will be able to recognize... -
CRISPR Guide
TypeGuide...joining (NHEJ) pathway and the less efficient but high-fidelity homology directed repair (HDR) pathway....also enhanced specificity by engineering Cas9 into high fidelity Cas9s (hfCas9) . There is currently no ..., which are about the same size as SaCas9, with higher fidelity than SpCas9. FrCas9 (from Faecalibaculum... downstream of the target site (termed left and right homology arms). The length of each homology arm ...available with multiple Cas9 variants, including high fidelity Cas9s, Cas9s with various PAM requirements...strands Prime editing and base editing both offer high efficiency and low indel formation compared to HDR...favor insertion in the T-RL direction, with the right homology end closest to the target site. Type V-... -
Adenovirus Guide
TypeGuide...simplicity of production, and higher titers. However, the risk of RCA formation is higher for first-generation ...rAdV vectors are also referred to as "gutless" or high capacity AdV (HCAdV) vectors. Here, all viral sequences...adenoviral backbone plasmid have matching left and right homology arms which facilitate homologous recombination...the shuttle plasmid, but this method results in a higher background of non-recombinant adenoviral plasmids... used for rAdV vector construction, due to their high gene transfer efficiency. Other serotypes like Ad26...immunogenicity. For example, Ad5/35 and Ad5/26 combine the high transduction efficiency of the Ad5 serotype with... particle. Vaccine Development rAdV vectors are highly immunogenic because they activate both innate and... -
Plan Your Experiment
TypeGuide...for a given experiment. In the table below, we highlight the most common categories of genetic manipulations...joining (NHEJ) at the cut site Cas9 or Cas9 nickase High-fidelity Cas enzymes increase specificity Dual-nickase...editors Editing efficiency can be low, but generally higher than HDR pegRNA optimization can be challenging...location. Positioning your chosen Cas enzyme at the right spot within the genome relies on proper gRNA design...design a donor DNA to have the appropriate left and right homology arms. See Addgene's HDR blog post for more...guides that target closer to the edit site yield higher editing efficiencies. The template sequences can...specificity, consider using an engineered Cas enzyme with higher fidelity. In addition to off-target activity, it... -
Adeno-associated virus (AAV) Guide
TypeGuide...episome which can concatemerize, producing high molecular weight structures that are maintained extrachromosomally...hybrid capsid derived from eight different serotypes. AAV-DJ displays a higher transduction efficiency in...on transgene expression and the expression of neighboring genes. rAAV episomes can develop chromatin-like...in vitro than any wild-type serotype, and a very high infectivity across a broad range of cell types in...Brain Connectivity Atlas and the Janelia Campus MouseLight project . rAAV vectors have also been developed...signals of cells (commonly neurons) in response to light. The short coding sequences of channelrhodopsins...wild-type). Another split AAV method which has shown higher efficiency in delivering large genes makes use ... -
Chemogenetics Guide
TypeGuide...opto-chemogenetic fusion proteins of a light-emitting luciferase and a light-sensing optogenetic element. When...neurotransmission. This, and the fact that GPCR ligands can have high potency, which is potentially important for dosing...off-target effects in vivo and the receptors exhibited high levels of constitutive activity, making them less...luciferin) is added, the luciferase enzyme generates light that activates the opsin. The opsin may excite or...illumination), making LMOs a versatile option. Coupling brighter bioluminescent modules, such as luciferase-fluorescent...example, a study in rats using an hM4Di receptor might consider an alternative DREADD ligand such as DCZ... chemogenetics by using physical and biological light sources for opsin activation. Proc Natl Acad Sci... -
Molecular Biology Reference
TypeGuide...insert expression (i.e., a strong promoter directs high expression, whereas weaker promoters can direct ...determines the type of plasmid and dictates how it might be used in the lab. Below are some common plasmid...particles, that can infect your target cells at a high efficiency. Addgene's expanding viral service offers...Next Generation Sequencing (NGS) comes in. NGS is a high-throughput, multi parallel sequencing platform that... Plasmids 101: 5 factors to help you choose the right cloning method Plasmids 101: Restriction Cloning... -
Cloning
TypeGuide...choosing the right cloning method for your research project . The following guide will highlight several of... -
Gamma-Retroviral Vector Guide
TypeGuide...for fast and efficient generation of large-scale, high-titer preps. This is thanks to established retroviral... on a case-by-case basis. Retroviruses carry a slightly elevated risk of integrating into proto-oncogenes...clinical-grade gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing... -
Science Guides
TypeGuide...immune system, antibodies are proteins capable of highly specific binding to other proteins. Antibody-based...technologies developed for optogenetics research utilize light to detect, measure, and control molecular signals... -
Lentiviral Vector Guide
TypeGuide...J., Bae, S., & Hur, J. K. (2021). CReVIS-Seq: A highly accurate and multiplexable method for genome-wide...Kochenderfer, J. N., Norberg, S. M., Hinrichs, C., Highfill, S. L., Somerville, R. P., Panch, S. R., Jin, ... -
Guide to Using Pooled Libraries
TypeGuide...additional hypothesis-directed experiments. As you might have guessed from their name, pooled libraries are... -
Promoters
TypeGuide...polymerase) to bind to a promoter. Transcription is tightly controlled in both bacteria and eukaryotes. Promoters... -
Which Fluorescence Microscopy Technique is Best for Me?
TypeBlog Post...limit of a light microscope. Due to the wave nature of light, an infinitely small point of light will blur... to provide high 3D spatial resolution while also avoiding the toxic effects of high light doses to the...fluorescence microscopy provides a high contrast image in which a bright signal overlays a perfectly black...placed to only allow light from the focal plane to reach the detector and all light from above or below...for example, red light instead of blue to excite a molecule of GFP. Because red light has approximately...approximately half the energy of blue light, two photons of red light are needed to excite GFP compared to only ...objective can a a high enough density of red-photons be establish for this to happen.Using red light has a second... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog Post...UV-Violet Dark/Green N/A (high) High 503/ 518 Padron Monomer Blue Dark/ Green N/A (high) Low 503/ 518 rsCherry...specific wavelength of light. PA-FPs can be activated from low fluorescent states to high fluorescence states...fluorescence, to a brighter fluorescent state by exposure to a specific wavelength of light. The switch or...fluorescence when illuminated with 488nm light. However, when UV-violet light is presented first, it causes irreversible...illuminated with violet light, both variants are photoconverted to forms that emit brighter red fluorescence....when exposed to blue light, which is less phototoxic to live tissue than the UV light used by other PA-FPs...State Activating Light Pre/ Post Color Contrast (Fold Change in Fluorescence) Brightness Excitation/ Emission... -
Viral Vectors 101: Optogenetic Tools
TypeBlog Post...with exquisite precision using only light. Optogenetics, or light activated tools, can control everything...an experimental optogenetic setup. A light source delivers light pulses to the subject/cells expressing... photoswitches. Light-gated ion channels directly open and close when exposed to light and change the ...changes with light stimulation to either activate or inhibit a specific receptor or pathway. Light-gated ion...turn on with a light-pulse at one wavelength and stay on until a second pulse of light, with a different...pathways with light. This is achieved by coupling endogenous signaling molecules with light-activated partners... intensities of light sources used to prevent activation of both tools with each light source used. ...