Methods and systems for nucleic acid synthesis
AssigneesReplay Therapeutics, Inc., Neochromosome Inc.
InventorsLeslie MITCHELL, Adrian WOOLFSON
Life of this patent
Estimating the term…
Abstract
The present disclosure provides methods, compounds, and systems for synthesizing a nucleic acid molecule. The methods may comprise spatially separating a set of barcoded oligonucleotides corresponding to a target nucleic acid molecule and performing a nucleic acid assembly using the oligonucleotides. A computer system coupled to a process control software program may be configured to apparatuses and configured to carry out the methods of the present disclosure.
Claims30
1-111. (canceled)
A method of generating a target polynucleotide, said method comprising:
- (a) providing a nucleic acid molecule comprising a stem-loop and a barcode sequence;
- (b) contacting a solid support with said nucleic acid molecule, wherein said solid support comprises a capture sequence complementary to said barcode sequence, thereby forming a capture complex comprising said capture sequence and said nucleic acid molecule;
- (c) separating said nucleic acid molecule from said capture complex;
- and
- (d) incubating said nucleic acid molecule with assembly reagents, thereby generating at least a portion of said target polynucleotide.
An oligonucleotide comprising a barcode sequence at a position, wherein said position:
- is within a 5′ localized stem loop of said oligonucleotide subsequence;
- is adjacent to a type IIS restriction site of said oligonucleotide;
- or
- is within a stem loop encoded near said 5′ stem loop of said oligonucleotide.
Dependencies are inferred from the claim text.