Patent ID: 11873374
Assignee: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Field: Measurement (Instruments)
Classification: CPC C  G | IPC C  G

Claim 9:
10. A method for in-situ sequencing of target nucleic acids present in a biological sample comprising the steps of:
a. attaching target nucleic acids present in the sample with a molecule linker or nucleic acid adapter;
b. permeating the sample with a monomer of Formula (A1):, and a monomer of Formula (B1):, to form a hydrogel according to claim 1 by non-radical polymerization and thereby forming a sample-hydrogel complex, wherein the small molecule linker or nucleic acid adaptor is attached both to the target nucleic acids present in the sample and to the hydrogel;
c. digesting proteins present in the sample;
d. expanding the complex to form a first enlarged sample;
e. re-embedding the first enlarged sample in a non-swellable material to form a re-embedded complex;
f. modifying the target nucleic acids or the nucleic acid adaptor to form a target nucleic acids or a nucleic acid adaptor; and
g. sequencing the nucleic acids present in the re-embedded complex;
wherein:
each n is an integer greater than or equal to 1;
each p is an integer greater than or equal to 1;
Z+ is a counter cation;
X and Y1 are each crosslinkable moieties, and
X and Y1 covalently crosslink to end-link the monomers.