Target C : Simple Hands-On Programming With Vis...
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Target C : Simple Hands-On Programming With Vis...
Figure 1. Common problems that arise with primers and 3-step PCR amplification of target DNA. (a) Self-annealing of primers resulting in formation of secondary hairpin loop structure. Note that primers do not always anneal at the extreme ends and may form smaller loop structures. (b) Primer annealing to each other, rather than the DNA template, creating primer dimers. Once the primers anneal to each other they will elongate to the primer ends. (c) PCR cycles generating a specific amplicon. Standard 3-step PCR cycling include denaturation of the template DNA, annealing of primers, and extension of the target DNA (amplicon) by DNA polymerase.
IPM is an ecosystem-based strategy that focuses on long-term prevention of pests or their damage through a combination of techniques such as biological control, habitat manipulation, modification of cultural practices, and use of resistant varieties. Pesticides are used only after monitoring indicates they are needed according to established guidelines, and treatments are made with the goal of removing only the target organism. Pest control materials are selected and applied in a manner that minimizes risks to human health, beneficial and nontarget organisms, and the environment.
Optical imaging offers great opportunities to monitor and analyze disease-related metabolites in live cells with a high spatiotemporal resolution1,2,3,4,5. Recent advances achieved in the development of fluorescence imaging techniques and biosensing systems allow for the precise and rapid detection of intracellular species for biomedical applications6,7,8,9. Ideally, fluorescence probes are designed to selectively recognize a target species in complicated biological systems. However, determination of a given analyte in a realistic biological milieu (such as in cells and in vivo) with fluorescence is easily interfered by the complexity of microenvironment (e.g., change of pH and salt strength, and the existence of structurally complicated biomacromolecules) and the inevitable background signals10,11,12,13. In particular, fluorescence probes that rely on the emission intensity change (fluorimetric) are prone to being suffered from these issues. To circumvent this problem, several elegant approaches have been proposed including the ratiometric sensing rationale (emission shift upon selectively recognizing an analyte), which is currently a method of choice for biosensing and bioimaging14,15,16.
The SP-Gal (Fig. 1a and Supplementary Fig. 18) and a control compound SP-PEG (Supplementary Fig. 19) with a polyethylene glycol (PEG), instead of Gal, were synthesized. SP is a popular photochromic molecule, which can be light-converted to the charge-separated zwitterionic MR structure after photoisomerization19, 25, 29. A variety of photochromic probes have been constructed based on SP since the zwitterionic isomer (MR) offers a coordination site for various analytes, such as ions and biomolecules30,31,32. In addition, photoisomerization of SP produces the MR isomer with an extended conjugated system, which can serve as a FRET acceptor to tune the fluorescence of closely coupled fluorophores33, 34. The double bond of the hemicyanine-like motif of MR is also prone to undergoing Michael addition reactions with nucleophiles, making possible the development of reaction-based probes35,36,37. Bearing these points in mind, the glycoprobes were synthesized through a simple coupling of naphthalimide with SP, producing SP-Gal in good yields. The fluorophore (naphthalimide) was coupled with D-galactose to target a specific transmembrane glycoprotein receptor, by a click reaction. The control compound (SP-PEG) was synthesized in a similar way.
The description of a programming language is usually split into the two components of syntax (form) and semantics (meaning), which are usually defined by a formal language. Some languages are defined by a specification document (for example,