• It is one of centerpieces of their SMS efforts, but presently it is hard to gauge process

• Because of the time and cost have been reduced, the error rates of third-generation sequencing (TGS) reach up to 4-11% and are relatively higher than second-generation sequencing (SGS)

• The choice of filters for fluorescent wavelength selection is also critical to the success or failure of experimental detection of FRET

• Contaminating direct excitation of the acceptor molecule can be accounted for using appropriate controls, but large amounts will make interpretation of the data more difficult

• Only those molecules that interact with one another will result in FRET. If large amounts of donor and acceptor molecules are present, but do not interact, the amount of FRET taking place would be quite low

• Dominique Francois Jean Arago, thefamous French astronomer repeated Oerstead's experiment and suggested the possibility of a telegraph together with Jacques Babinet
• Faraday pictured lines of force as the mechanism by which electrical and magnetic substances interact with themselves and with each other
• James Clearn Maxwell created a compeleted mathematical representation of Faraday's descriptions of electricity and mangnetism
• First theoretical attempts explaining FRET were applications of this classical EM theory

• Then, he introduced the notion of dispalcement current and furnished theoretical setting to predict electromagnetic radiation
• Hertz carried out the critical experiment in 1888, and published the theory to explain the EM fields surrrounding his electric oscilliator in 1889
• First experiment were conducted by Hertz by producing high frequency repetitive sparks in an air gap of a primary oscillating circuit
• His theoretical description the electromagnetic disturbance in the near field where the electromagnetic energy escapes and is carried away as radiation with tranverse oscillations
  

• Maxwell's theory reaches very unsatisfactory state, Planck solved this by introducing the quantum concept
• Bohr's theory of the atom be introduced
• Quantum mechanical theoty to explain the transfer of energy between atoms at longer distances compared to the collisional radii was proposed by Kallmann and London20
• J. Perrin hypothenized that the transfer of the excitation energy could hop from one molecule to the other through interactons between oscillating dipoles of closely speaced molecules
• Oppenheimer with Arnold explained about photosythesis
• Forster then derive a quantitative theory of non-radiative energy transfer that still use until this day
• He also furnished the clear and explicit to experiment

• Donor fluorophore absorb the energy due to the excitation of incident light. The donor will transfer the excitation energy to a nerby chromorphore, the acceptor through long range dipole-dipole intermolecular coupling
• The energy transfer will manifests itself through decrease or quenching of the donor fluorenscence and a reduction of excited state lifetime also acompanied by the rate changes in acceptor fluorenscence
• The important criteria in FRET is the fluorenscence emission spectrum of the donor molecule must overlap the absorption spectrum of the acceptor,The donor and acceptor must be in the close proximity to one another, The transition dipole proentations of the donor and acceptor must be in parallel state towards each other and lastly, the fluorenscen lifetime of the donor molecule must be of sufficient duration to allow the process of FRET to occur
  

• The two involved fluophores must be positioned within a range of 1 to 10 nanometers of each other
• The distance dependence of the resonance energy transfer process is the primarily basis in investigation of molecular interaction
• In living cell studies that involving donor and acceptor fluorophores, resonance energy transfer will occur only between molecules that are close enough to interact biologically with one another