Compositions and methods for intramolecular nucleic acid rearrangement
Aspects of the present invention are drawn to processes for moving a region of interest in a polynucleotide from a first position to a second position with regard to a domain within the polynucleotide, also referred to as a “reflex method”. In certain embodiments, the reflex method results in moving a region of interest into functional proximity to specific domain elements present in the polynucleotide (e.g., primer sites and/or MID). Compositions, kits and systems that find use in carrying out the reflex processes described herein are also provided.
1. A composition comprising a mixture of:
a) a first plurality of different segments of a first nucleic acid molecule of a contiguous region in genomic DNA, wherein said contiguous region in genomic DNA is represented by a plurality of nucleic acid molecules, and wherein each of said first plurality of segments is appended to a first multiplex identifier (MID) sequence; and
b) a second plurality of different segments of a second nucleic acid molecule of said contiguous region, wherein each of said second plurality of segments is appended to a second MID sequence and said first MID sequence and said second MID sequence are different,
wherein, in said composition, all of the segments of each of the nucleic acid molecules of the contiguous region comprise a unique MID sequence that uniquely identifies each of the nucleic acid molecules.
2. The composition of claim 1 , wherein said genomic DNA is mammalian genomic DNA.
3. The composition of claim 2 , wherein said mammalian genomic DNA is human genomic DNA.
4. The composition of claim 1 , wherein the segments of a) and b) are in the range of 100 to 5,000 bp in length.
5. The composition of claim 1 , wherein the segments of a) and b) are 400 bp or less in length.
6. The composition of claim 1 , wherein:
the different segments of said first nucleic acid molecule are distributed along said first nucleic acid molecule; and
the different segments of said second nucleic acid molecule are distributed along said second nucleic acid molecule.
7. The composition of claim 6 , wherein:
the different segments of said first nucleic acid molecule are distributed along said first nucleic acid molecule at intervals in the range of 250 bp to 5 kb; and
the different segments of said second nucleic acid molecule are distributed along said second nucleic acid molecule at intervals in the range of 250 bp to 5 kb.
8. The composition of claim 1 , wherein:
substantially all of said first nucleic acid molecule of said contiguous region is represented in said first plurality of segments; and
substantially all of said second nucleic acid molecule of said contiguous region is represented in said second plurality of segments.
9. The composition of claim 1 , wherein said contiguous region is up to 30 kb in length.
10. The composition of claim 1 , wherein said genomic DNA is genomic DNA from a virus or a bacterium.
11. The composition of claim 1 , further comprising a third plurality of different segments of a third nucleic acid molecule of said contiguous region, wherein each of said third plurality of segments is appended to a third MID sequence and said third MID sequence is different to said first and second MID sequences.