Phased genome sequencing

摘要

The present disclosure provides methods for determining phased nucleic acid sequence for a single chromosome of interest and/or a single chromosomal fragment of interest. The present disclosure also provides methods for determining phased nucleic acid sequence for a plurality of single chromosomes of interest and/or a plurality of single chromosomal fragments of interest. The plurality of single chromosomes of interest may be of one chromosome type or of two or more chromosome types. The present disclosure also provides a method for isolating a plurality of chromosomal fragments of a specified size range, where the chromosomal fragments are from one or more specified regions of the genome. The plurality of chromosomal fragments may be separated into single chromosomal fragments and sequenced to provide phased nucleic acid sequence for the single chromosomal fragments. Alternatively, the plurality of chromosomal fragments may be sequenced together to provide unphased nucleic acid sequence for the chromosomal fragments.

主权项

1. A method comprising:
fragmenting a chromosome into chromosomal fragments; contacting chromosomal fragments with a plurality of oligonucleotide probes, wherein the oligonucleotide probes contain a fluorescent signal, wherein the oligonucleotide probes bind to a nucleic acid comprising a nucleic acid sequence of interest, isolating chromosomal fragments based on the fluorescent signal by using a fluorescence activated cell sorter, wherein the isolated chromosomal fragments associated with the fluorescent signal are separated from chromosomal fragments not associated with the fluorescent signal, wherein the isolating comprises collecting the isolated chromosomal fragments associated with the signal into separate containers each container containing no more than one chromosomal fragment; sequencing the isolated fragments associated with the fluorescent signal to obtain sequence of the chromosomal fragment containing the nucleic acid sequence of interest.