Systems and methods for epigenetic sequencing
The present invention generally relates to microfluidics and/or epigenetic sequencing. In one set of embodiments, cells contained within a plurality of microfluidic droplets are lysed and the DNA (e.g., from nucleosomes) within the droplets are labeled, e.g., with adapters containing an identification sequence. The adapters may also contain other sequences, e.g., restriction sites, primer sites, etc., to assist with later analysis. After labeling with adapters, the DNA from the different cells may be combined and analyzed, e.g., to determine epigenetic information about the cells. For example, the DNA may be separated on the basis of certain modifications (e.g., methylation), and the DNA from the separated nucleosomes may be sequenced using techniques such as chromatin immunoprecipitation (“ChIP”). In some cases, the DNA sequences may also be aligned with genomes, e.g., to determine which portions of the genome were epigenetically modified, e.g., via methylation.
1. A method, comprising:
providing a plurality of droplets containing cells, the droplets formed of a first liquid contained within a second fluid;
lysing the cells contained within the droplets to produce cell lysate within the droplets;
exposing at least some of the cell lysate contained within the droplets to a nuclease to produce a plurality of sequences within the droplets, wherein the nuclease is a non-nucleosome-cleaving nuclease;
attaching at least some of the sequences to an adapter, the adapter comprising an identification sequence and a restriction site, wherein the identification sequences allow identification of the cells within the plurality of droplets, and wherein the adapter is palindromic;
sequencing at least some of the sequences.
2. The method of claim 1 , wherein the plurality of sequences comprise DNA sequences.
3. The method of claim 1 , wherein the plurality of sequences comprise RNA sequences.
4. The method of claim 1 , wherein the identification sequences comprise 8 nucleotides.
5. The method of claim 1 , wherein a plurality of adapters are used, comprising a plurality of different identification sequences each having the same length, and a substantially identical restriction site.
6. The method of claim 1 , wherein the adapters, prior to attachment, are double-stranded.
7. The method of claim 1 , wherein the adapter further comprises a primer.
8. The method of claim 1 , wherein sequencing the sequences comprises performing chromatin immunoprecipitation (ChIP) sequencing on the sequences.
9. The method of claim 1 , further comprising amplifying at least some of the sequences.
10. The method of claim 1 , wherein at least some of the cells arise from tissue.
11. The method of claim 1 , wherein sequences originating from the same cell contain identical identification sequences, and sequences originating from different cells contain different identification sequences.
12. The method of claim 1 , wherein at least some of the sequences are ligated to an adapter.