... difficult to quantify accurately, but can be especially helpful to generate more signal from low abundance...the multicolor overlay; this helps all viewers, especially if you have some complex patterns with several...

...recombinase expression may be replaced by sensors that correspond to the desired inputs. Such sensors do not necessarily...described in Nielsen et al (3) and Lou et al (4). Despite choosing proper register parts and designs, we ...

...allows you to easily orient your gene insert with respect to the promoter. For example, let’s say your plasmid...being made (post or pre run in the table above, respectively). Some of the above stains require you to visualize...

...validation techniques for academia, industry, and especially the clinic. Popular validation assays are insufficient...search the whole genome for off-target sites irrespective of in silico prediction. These techniques differ...

...important to test it in the brain region of interest, especially when using DIO/FLEx vectors for off-target expression...relatively low recombination rate (0.1% and 0.13% respectively) still required a 200-300 fold dilution to minimize...

... lab Undergraduate student Roles and responsibilities can vary "Lab assistants" usually wash glassware... my experience in the lab? Ask your questions—especially if it’s scary. I’ve been afraid to ask “dumb ...

...considerations For changes 5’ to the PAM, sequences corresponding to the same strand as the PAM produced a higher...efficiency, while for edits 3’ to the PAM sequences corresponding to the opposite strand were better (Farboud ...

...sequences at the intended target site. With the widespread adoption of CRISPR/Cas9 technology and rapid ...researchers from generating clonal knock-in lines, especially for ambitious projects that require epitope tagging...

...lower than the energy of the absorbed photon. Correspondingly, wavelengths of emitted photons are higher ...FRET).   Figure 1: Emission spectra and respective colors of a few commonly used fluorophores. Each...

...through the excitation light path. Each hole corresponds to a different location in the sample. As more...step that reassigns out of focus light to its respective focal plane. In sufficiently thin samples, deconvolution...

... of a pathway to inhibit to block the cellular response to a particular ligand (such as when a component...pathway. For example, one could study the cellular response to a cytokine known to cause the up-regulation...

...mastered this technique so it might be worth a try, especially since the potential rewards could be substantial...) distance at which 50% FRET occurs. b Range corresponds to calculated D/A distances for observed FRET...

...bricks” to be added one by one. This approach is especially useful for combinatorial analyses. For example... L-arabinose and anhydrotetracycline (aTet), respectively. pMAZ-SK contains an aTet-inducible gRNA and...

...chemogenetic tools are unresponsive to native ligands and are engineered to respond to small molecules that...PSAM that responded to a clinically approved drug, the PSAM domain was engineered to respond specifically... development of mutated receptors that while unresponsive to endogenous ligands, could be activated by...Gs-DREADD, rM3D, that was created by replacing the corresponding intracellular region of a turkey erythrocyte...Muscarinic-receptor based DREADDs were engineered to respond to nM concentrations of clozapine N ‐oxide (CNO... shown to be an agonist against hM4Di, and is especially attractive for use in translational studies because...of a suite of muscarinic-based DREADDs that all respond to the same ligands was that bidirectional experiments...

...labelled with a green or violet, fluorescent dye, respectively. WT and KO cells were mixed in a 1:1 ratio and...the indicated Synaptotagmin-1 antibodies and corresponding Multi-rAb CoraLite® Plus 647 secondary antibodies...

...caused by not having the time to assess public response; and, it is the stage I think we are currently...effort from the community in order to succeed. With respect to de novo sgRNA design, I again separate it into...

... to de-differentiate. However it’s difficult, especially as a newbie reprogrammer, to decide which method...reprogramming transgenes and the up-regulation of corresponding endogenous pluripotency genes. Retroviral vectors...

... peak indicating 2 amplicons. Image credits (respectively): Zuzanna K. Filutowska; Selma, 2019; Trampuz...you’re using a different brand of qPCR machine, especially if you’re using a rapid-cycle machine such as...

...animal, which then mounts an immune response to it. Once the immune response has developed, the antibodies ...Antibodies, produced as part of the immune system’s response to foreign bodies, are the most commonly used ...Figure 1. The two arms of the Y structure are responsible for binding the antigen, and are called the antigen...bind to a specific antigen, triggering an immune response. The stem of the Y structure and the lower part...isotype of the antibody. B cells, the immune cell responsible for producing antibodies, can change the constant...binding site. In B cells, the genetic region responsible for the variable region goes through a randomized...Different isotypes play different roles in the immune response, and can be used to increase flexibility or specificity...

...primary antibodies can be more difficult to source, especially if you need a specific conjugate, and changing...detection quickly (~1 minute), and have a linear response over a broad dynamic range that typically spans...