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US 2023/0054869 A1Published application
Methods and Compositions Employing Blocked Primers
AssigneeTakara Bio USA, Inc.
InventorsThomas D. Schaal, Jude Menlo Dunne, Maithreyan Srinivasan, Alain Mir
Life of this patent
Estimating the term…
Abstract
The present disclosure provides methods, compositions, and systems employing blocked primers. Aspects of the disclosure include providing a blocked primer reaction mixture that includes a blocked primer and a template nucleic acid component from a single cell; unblocking the blocked primer to produce an active primer reaction mixture and subjecting the activated primer reaction mixture to primer extension conditions, such as nucleic acid implication conditions.
Claims30
1-15. (canceled)
A method comprising:
- (a) combining a blocked primer comprising a light labile blocking moiety with a template nucleic acid component to produce a blocked primer reaction mixture;
- (b) unblocking the blocked primer of the blocked primer reaction mixture by exposing the blocked primer reaction mixture to light sufficient to remove the blocking moiety to produce an activated primer reaction mixture;
- and
- (c) subjecting the activated primer reaction mixture to template dependent primer extension reaction conditions.
1 more element
A method comprising:
- (a) combining a blocked primer comprising a thermally labile blocking moiety with a template nucleic acid component to produce a blocked primer reaction mixture;
- (b) unblocking the blocked primer of the blocked primer reaction mixture by heating the blocked primer reaction mixture to a temperature sufficient to remove the thermally labile blocking moiety to produce an activated primer reaction mixture;
- and
- (c) subjecting the activated primer reaction mixture to template dependent primer extension reaction conditions.
1 more element
A method comprising:
- (a) combining a blocked primer comprising an enzymatically labile blocking moiety with a template nucleic acid component to produce a blocked primer reaction mixture;
- (b) adding an enzyme to the blocked primer reaction mixture to enzymatically remove the enzymatically labile blocking moiety of the blocked primer, thereby unblocking the blocked primer of the blocked primer reaction mixture to produce an activated primer reaction mixture;
- and
- (c) subjecting the activated primer reaction mixture to template dependent primer extension reaction conditions.
1 more element
44-47. (canceled)
Dependencies are inferred from the claim text.