We narrowed to 586 results for: cel.2;
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TypeBlog Post...Table 2, Figure 5) for a hypothetical experiment in the lab. Let's say I am culturing HEK 293T cells in ... weak. Table 2: A simple flow panel to detect live, transduced, and CD45-expressing cells. Marker .... To understand why that is, let's look at Figure 2. The blue laser excites at a wavelength of 488 nm ... of PE's excitation spectrum (dotted line, Figure 2). The yellow/green laser, however, excites at 561 ... stronger — or brighter — signal. Figure 2: Excitation and emission spectra for PE. The excitation...efficiency (eGFP expression). The first item in Table 2 is a standard and should always be included—the live...from the other fluorophores to avoid bleeding and 2) BV421 is a very bright fluorophore, which makes it...
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Genome-wide Screening Using CRISPR
TypeBlog Post...plasmid, or a 2-plasmid system in which Cas9 must be delivered separately. Mutant cells are enriched ...diseases causing uncontrolled cell growth, such as cancer. Figure 2: The CRISPR library must be ...targeting a specific gene within the genome (see Fig. 2). Individual gRNAs are designed in silico using publicly...mouse, human, fly, E. coli, and T. gondii genes. 2) What genetic modification are you trying to make?...CRISPR libraries is a multi-step process (see Fig. 2). In most cases, CRISPR libraries are provided at ...heterogeneous population of mutant cells, with each cell or set of cells containing a mutation in a different...fluorescence-based cell sorting and screened for a particular phenotype. For example, mutant cells can be used... -
A Primer on Optogenetics: Introduction and Opsin Delivery
TypeBlog Post...: PMC3337962. 8. Zhao S, et al. Cell type-specific channelrhodopsin-2 transgenic mice for optogenetic ...commonly used in optogenetics: 1) electrophysiology and 2) behavioral experiments. I’m not an electrophysiologist...choice in the brain region you wish to stimulate 2) Fabrication and implantation of the optical fiber...thoroughly elsewhere (here and here for instance) [2], [3]. Karl Deisseroth of Stanford University , one...reviews for a discussion of the different variants [1, 2, 4]. And, of course, the best way to learn about the...monitor expression at various time points (e.g. every 1-2 weeks over 8 weeks) in order to determine the optimal...(4):251-66. doi: 10.1038/nrn3171. PMID: 22430017. 2. Fenno L, et al. The development and application of... -
Luciferase Plasmid Collection
TypeCollection...and Venus fluorophore to Troponin C Ca 2+ binding domain. Nano-lanterns , Enhanced Nano-lanterns , and...transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623-2) Firefly PGK Lentiviral expression of firefly luciferase...Expression of firefly luciferase from a cell-free mammalian cell lysate Marcel Bruchez 108542 pLenti-EF1a-Luciferase-IRES-Blast-WPRE...not secreted, so cells must be lysed or the substrate must be delivered intracellularly in order to measure... viral infection efficiency, to study changes in cell physiology, or as a reference level of expression...interactions , protein dimerization , endosomal escape of cell-penetrating peptides , and more. nLuc/cLuc for split...Resonance Energy Transfer (BRET) CalFlux VNT : An intracellular calcium sensor created by fusing NanoLuc® and... -
Five Popular Model Organisms, Part 2
TypeBlog Post... H. J. (2009). Rats! Disease Models & Mechanisms, 2(5–6), 206–210. https://doi.org/10.1242/dmm.002733 ...are often used to study vertebrate embryology and cell development. One major resource for frog research...Organism to Study and Model Human Genetic Disease. Cells Tissues Organs, 205(5–6), 303–313. https://doi.org... -
Viral Vectors 101: Optogenetic Tools
TypeBlog Post...Optogenetic control of intracellular signaling pathways. Trends in Biotechnology, 33(2), 92–100. https://doi.org...for controlling neuronal activity, channelrhodopsin-2 (ChR2), a blue light-activated cation channel, from...used in tandem with the opsin. Figure 2: Diagram of excitatory (ChR2) activation by light ...) domains, phytochrome B (PhyB), and cryptochrome 2 (CRY2) (Zhang & Cui, 2015). When stimulated with light...comparative analysis of microbial opsins. Nature Methods, 9(2), 159–172. https://doi.org/10.1038/nmeth.1808 References...comparative analysis of microbial opsins. Nature Methods, 9(2), 159–172. https://doi.org/10.1038/nmeth.1808 Nagel...Hegemann, P., & Bamberg, E. (2003). Channelrhodopsin-2, a directly light-gated cation-selective membrane ... -
Brain Initiative Collection
TypeCollection...Gordon Fishell 83899-AAV1 pAAV-mDlx-GCaMP6f-Fishell-2 GCaMP6f expression in forebrain GABA-ergic interneurons...Gordon Fishell 83899-AAV9 pAAV-mDlx-GCaMP6f-Fishell-2 GCaMP6f expression in forebrain GABA-ergic interneurons...Gordon Fishell 83899-AAVrg pAAV-mDlx-GCaMP6f-Fishell-2 GCaMP6f expression in forebrain GABA-ergic interneurons...real-time interrogation and monitoring of brain cells and circuits during behavior. Long desired by researchers...A somatic channelrhodopsin, which enables single cell, single millisecond resolution optogenetics. Human...A somatic channelrhodopsin, which enables single cell, single millisecond resolution optogenetics. Cre-dependent...genetically encoded dopamine sensor RdLight1 in mammalian cells. Lin Tian 125708-AAV9 pAAV-hSynapsin1-RdLight1 Expresses... -
Plasmids 101: Dimers and Multimers
TypeBlog Post...identification. Fig. 2: Gel image of Plasmid 61564: pBAMD1-2 (~4.7 kb) digested with SpeI and...to the formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674(76)90134... at ~3.6 kb and ~1.1 kb (lane 3). Uncut DNA (lane 2) has a band at <3 kb, which corresponds to the supercoiled...dimers in Escherichia coli K-12. Plasmid. 1989 Mar;21(2):138-41. doi: 10.1016/0147-619x(89)90057-7. PMID: ...Plasmids and Chromosomes. Microbiol Spectr. 2014 Oct;2(5). doi: 10.1128/microbiolspec.PLAS-0025-2014. PMID...Behavior of Plasmid DNA. Microbiol Spectr. 2015 Apr;3(2):10.1128/microbiolspec.PLAS-0036-2014. doi: 10.1128...at lower copy numbers within bacterial cells, and bacterial cells with multimers grow more slowly compared... -
Flow Cytometry Readouts: Yes, No, and Everything in Between
TypeBlog Post...low expression, and everything in between (Figure 2). These smears tend to occur when you are looking ...on your experiment. You can see from Figure 2 that determining where to place your gate based on the dot... dot plot would be complicated. Figure 2: (A) a smeary dot plot showing a range of antibody readouts...plot in Figure 4 compared to the dot plot in Figure 2. Quantification using MFI makes it easier to identify...percentage of total live cells are GFP+. In this plot, with adjusted voltage, cells that did not receive ...would serve you well. Percentage of cells in a parent gate (all live cells for example) that are positive ...example, you have introduced a GFP protein into your cells (visualized via the FITC channel) and you are trying... -
CRISPR 101: Targeting RNA with Cas13a (C2c2)
TypeBlog Post...question: C2c1, C2c2 and C2c3 (short for Class 2, candidate 1, 2, or 3). C2c1 and C2c3 are related to Cpf1,...characterization of diverse class 2 CRISPR-Cas systems." Molecular cell 60.3 (2015): 385-397. PubMed PMID...however cutting efficiency of Cas13a is reduced when 2 mismatches are present. The protospacer flanking sequence...PMC5127784. Find plasmids from this paper at Addgene. 2. East-Seletsky, Alexandra, et al. "Two distinct RNase...thought to activate programmed cell death or a dormant state for bacterial cells that have been infected with...bacterial cells, LwaCas13a does not display nonspecific RNA cleavage activity in eukaryotic cells. They arrived...target RNAs, although its usefulness in bacterial cells will likely be limited by the propensity of Cas13a...