Single-molecule sequencing

Smith T. Whole genome variation analysis using single molecule sequencing. Drug Discov Today Targets 2004 3 112-116. 

Non-SELEX is an attractive alternative to SELEX, due to increased speed and suitability for easy automation. Non-SELEX leads to a final pool with all sequences being unique. The analysis of such sequences would be much more efficient if single-molecule sequencing were used. In this respect, the current effort in the development of single-molecule sequencing technologies is quite encouraging (Bayley, 2006). 

Brackmann S and Lobermann S (2002) A further step towards single-molecule sequencing Escherichia coli exonuclease III degrades DNA that is fluorescently labelled at each base pair. Angewante Chemie International Edition English 41 3215-3217. 

Enzyme microreactors have also received wide application not only in the analysis but also in the synthesis of DNA. A nanoscale platform for real-time single-molecule sequencing by recording the DNA synthesis process by an immobilized DNA polymerase has been developed and has been covered by US patent application number 7,329,492 [62]. The process relies on fluorescence resonance energy transfer (FRET) between the fluorescence receptor moiety on the dNTPs and the fluorescent dye attached to the donor DNA that emits an enhanced signal upon nucleotide incorporation. Immobilization of polymerase enables increased sensitivity detection as the DNA strand extends imattached. 

Nikiforov, X Beechem, J. Methods and apparatus for single molecule sequencing using energy transfer detection. PCX Int. Appl. WO 2010111674, 2010. 

Kapanidis, A. Polymerase-based single molecule sequencing. PCT Int. Appl. WO 2009056831,... 

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