We narrowed to 988 results for: igh

... available with multiple Cas9 variants including high fidelity Cas9’s. Further advancements have been ...

...the challenge of trying to figure out what system might work best for your particular purposes. We’re here...than others. In today’s post, we will focus on a high-level comparison of popular cloning methodologies...results. If you’re in a hurry, Golden Gate cloning might be for you. This methodology relies on versatile... in as little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you ...fragments in an order determined by their homology. You might also consider FastCloning, first described by Chaokun...approach should you use if the budget is particularly tight? Most cloning processes will require a PCR reaction... job done, provided that you’ve already got the right enzymes laying around the lab. If not, you’ll be...

...and wrap the column with blue LED lights. When you switch off the lights, OptoNBs bind YFP and non-specific... the blue lights. Which option do you choose?  Option #2 may sound like fiction, but light-controlled...dark-induced dark-induced Loop 6 blue-light induced dark-induced blue-light induced Table 1: Summary of the... effect of AsLOV2 insertion site on light-induced binding.  Light controlled protein purification with...while cycling between dark and blue light which revealed a blue light-dependent shift in mCherry fluorescent...cells: light-based control of signaling pathways. This is particularly exciting because using light to control...binding of their target protein is altered by blue light illumination. This ability to reversibly bind and...

...engineering of fusion proteins is relatively straightforward. However, generating a fusion while keeping...discuss strategies to generate fusion proteins and highlight some aspects of their design.  Protein size and...situation Next to size, potential homodimerization and higher order oligomerization of fluorescent proteins is... the fluorescent tag should operate as an inert light emitting module, any tendency of GFP to homodimerize...fluorescent protein characteristics such as the brightness, photostability, maturation time, and acid sensitivity...termini of your protein of interest The most straightforward fusion protein is made by connecting the N-...the fluorescent protein-POI fusions, we generate straight fusions, without any linker. When both orientations...

...conducted in Professor Cepko's lab and to get some insight into her management and mentoring styles. If you'd...

...Gly-Ser in-frame with prefix start codon; Employs high-efficiency enzymes. Prohibited Restriction Sites...

... focused on three different levels of assembly: High efficiency and accurate single-insert assembly Intermediate...destination plasmid. Antibiotic selection allowed for high throughput cloning efficiency measurements. We performed...confirmed the insertion of the Lambda insert at the same high frequencies as the antibiotic selection markers....lacZ cassette assembly was easily achievable with high levels of transformants and low backgrounds – so...Using these 4 bp overhang observations, accurate “high-fidelity” junction sets for both the 12- and 24-...complex assemblies were performed. It was found that high efficiencies and accurate assembly levels were indeed...in the number of transformants while maintaining high levels of correct assemblies) translate to faster...

... (blue light-inducible AraC dimers in E. coli) that turns expression on and off with blue light.   Inspired...domain with VVD, a light-responsive domain that dimerizes upon exposure to blue light (Fig. 1). Another...VenturaIf you follow Barbara Di Ventura on Twitter, you might have seen the video of her singing about the paper...Another blue light-inducible dimerization domain, Vaucheria frigida aureochrome1 (VfAu1), led to functional...most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize and activates...under the pBAD promoter, can be controlled with light without the need of any cloning.     What advantages...BLADE activates transcription with the flick of a light switch and shuts off just as quickly. The original...

... in bacterial genome editing is not always straightforward. Read our CRISPR Methods for Bacterial Genome...

...This is where FLEx enters the scene. FLEx: A light when night is about you! The FLEx (for flip-excision)...optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford University, led by...ago with the discovery of microbial rhodopsins, light-sensitive proteins that absorb photons for energy...maintain membrane potential in response to visible light (Zhang et al., 2011). What do microbial rhodopsins...2000). Most neurons in the brain are not naturally light-sensitive, so the selective expression of opsin ...Optogenetic neuromodulation is made possible by the light: when hit with the correct wavelength, microbial..., 2011; Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger...

... The p53 name arose from its observed molecular weight (53 kDa) when run on an SDS-PAGE gel. Cancer Impact...

...hundreds of in-stock preps that are ready to ship right away, and our Packaged on Request service gives ...

... Casanova, J. E., Cherfils, J., Dacks, J. B., Lambright, D. G., Lee, F. S., Randazzo, P. A., Santy, L....

...showed that improved specificity of Cas14 enables high-fidelity SNP genotyping. The development of  Cas14...

...Scientist. I used to spend my days (and often my nights :D) engineering E. coli to produce biofuels in ... screaming from the lab like the cartoon on the right?!? I would love to tell such a dramatic story, but...This may come as somewhat of surprise to many of my high school friends. In my teenage years I was super ...can involve walking alongside it and giving it a high five. While screaming can be therapeutic and I’ve...

...erythroid 2 like 2 NFKB1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 PAK PAK1 ...

...called SARS-CoV-2. In 2020, scientists worked at lightning speed to elucidate as much as they could about...

...We provide high quality AAV preps from select plasmids in the repository. Browse our control AAV collection...

...vector serves as a backbone to clone the left and right homology arms. More details on how to use this plasmid...

...is not compatible with other plasmids containing high copy ColE1/PUC-like origins. Therefore, when choosing...

...Looger 106174 pAAV.hSynapsin.SF-iGluSnFR.A184S Syn Tight affinity glutamate sensor Glutamate sensor Loren...

... Lin28A to somatic cells from separate plasmids Highly Efficient Reprogramming to Pluripotency and Directed...

...that it is responsive to specific stimuli, such as light. On a broader scale, the expanded genetic code can...

...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...

...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...

...coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing)... 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...

...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...green-yellow light. Arch variants have been developed with the following enhancements: Increased light sensitivity...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...

...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...

...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-...

...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...