The patent's assignee for patent number 8785211 is
News editors obtained the following quote from the background information supplied by the inventors: "The current method for sequencing DNA involves a number of costly reagents such as fluorescent ddXTPs, dXTPs, primers and polymerase. This method requires sophisticated equipment, which needs to be operated by a qualified technician. Also, this method is limited to sequences of less than one thousand nucleotides in length.
"Other sequencing methods have been considered in order to reduce cost, simplify the method, and allow sequencing to take place out of the lab. Cycle extension, polymerase reading, exonuclease sequencing, and DNA micro-arrays are methods that have been considered (Braslaysky, I.,
"One potential method of sequencing DNA is based on threading a single strand of DNA through a nanopore and identifying its sequence from the variation in the ionic current flowing through the pore as the strand is threaded (Kasianowicz, J. J.,
"Stochastic sensing involves placing a nanometer sized pore in an insulating lipid bilayer membrane and measuring the ionic transport through the pore. When an analyte interacts with a binding site within the pore, a change in the ionic current is detected (Braha, O.,
As a supplement to the background information on this patent, NewsRx correspondents also obtained the inventors' summary information for this patent: "It has been surprisingly demonstrated that individual nucleotides can be identified at the single molecule level from their current amplitude when they interact with a transmembrane pore. Hence, stochastic sensing may be used to identify individual nucleotides and to sequence nucleic acid sequences via exonuclease sequencing.
"Accordingly, the invention provides a method of identifying an individual nucleotide, comprising: (a) contacting the nucleotide with a transmembrane protein pore so that the nucleotide interacts with the pore; and (b) measuring the current passing through the pore during the interaction and thereby determining the identity of the nucleotide.
"The invention further provides: a method of sequencing a target nucleic acid sequence, comprising: (a) digesting an individual nucleotide from one end of the target sequence using a processive exonuclease; (b) contacting the nucleotide with a transmembrane protein pore so that the nucleotide interacts with the pore; © measuring the current passing through the pore during the interaction and thereby determining the identity of the nucleotide; and (d) repeating steps (a) to © at the same end of the nucleic acid sequence and thereby determining the sequence of the nucleic acid; and a kit for sequencing a nucleic acid, comprising: a cyclodextrin; and a processive exonuclease.
"The method of sequencing of the invention is a rapid and simple DNA sequencing method at the single molecule level. It is also a cheap method of sequencing DNA because it does not involve the use of expensive reagents, such as fluorophores."
For additional information on this patent, see: Bayley, Hagan; Astier, Yann; Braha, Orit. Methods Using Pores. U.S. Patent Number 8785211, filed
Keywords for this news article include: Esterases, Polymerase,
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