Alternative nucleic acid sequencing methods
View Patent ↗Embodiments are provided that provide for parallel sequencing of nucleic acid segments. In some embodiments, a single sequence is sequenced by at least two different sequencing techniques and the results compared, allowing for deficiencies or strengths of one technique to be complemented by the second technique.
1. A method for sequencing a polynucleotide in a preparation by a sequencing instrument, wherein the sequencing instrument comprises a processor, said method comprising:
obtaining a polynucleotide preparation comprising the polynucleotide, comprising amplifying the polynucleotide in a semisolid support,
determining, by the sequencing instrument, a first apparent sequence of a first region of a polynucleotide by subjecting a sample of the polynucleotide preparation to a first type of sequencing technique;
simultaneously determining, by the sequencing instrument, a second apparent sequence of a second region of the polynucleotide that overlaps with the first region, by subjecting a sample of the same polynucleotide preparation to a second different type of sequencing technique,
wherein the first and second sequencing techniques are selected from the group consisting of Maxam-Gilbert sequencing, chain termination methods, dye termination methods, sequencing using reversible terminators, sequencing of nucleic acid by pyrophosphate detection, sequencing by ligation, closed complex single molecule sequencing, nanoscale fluidic sequencing, and force spectroscopy platform sequencing;
identifying, by the processor, any discrepancies between the first apparent sequence and the second apparent sequence; and
selecting, by the processor, the least error prone sequence from the first apparent sequence and the second apparent sequence based upon the likelihood of an error or error rate in one sequencing technique compared to the other sequencing technique.
2. The method of claim 1 , wherein the polynucleotide is immobilized on a solid support.
3. The method of claim 2 , wherein the solid support is a bead.
4. The method of claim 1 , wherein the polynucleotide is a single molecule.
5. The method of claim 1 , wherein the polynucleotide is an amplified clone.
6. The method of claim 5 , wherein the amplified clone is produced by PCR.
7. The method of claim 6 , wherein the PCR is emulsion PCR.
8. The method of claim 5 , wherein the amplification takes place on a solid support.
9. The method of claim 5 , wherein the amplified clone is present on an array.