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US 2023/0013384 A1Published application
High Throughput Reaction Assembly
AssigneeNew England Biolabs, Inc.
InventorsGuoping Ren, Yan Xu, Dong Ma, Nicole Nichols
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Abstract
Provided herein is a reverse transcriptase mixture comprising a reverse transcriptase and a colored dye at a concentration in the range of 0.003%-1% (v/w). The colored dye may be visually observed during transfer of the mix from one vessel to another and addition of the mix to another mix can be confirmed by eye by observing the colored dye.
Claims7
-16. (canceled)
A method comprising:
- (a) combining:
- (i) a supermix that comprises a reverse transcriptase, one or more primers, xylene cyanol and/or orange G and at a concentration of that is visible to the naked eye but inhibits Taq polymerase, and no Taq polymerase;
- with
- (ii) RNA, to produce a cDNA synthesis mix;
- (b) incubating the cDNA synthesis mix for a period of time to produce cDNA;
- (c) combining an aliquot of the product of step (b) with receiving mix that comprises a Taq polymerase and a primer pair to produce a PCR mix, wherein the xylene cyanol or orange G is diluted in concentration by at least 5-fold by the receiving mix;
- (d) thermocycling the PCR mix of step (c);
- and
- (e) detecting a reaction product of step (d).
6 more elements
Dependencies are inferred from the claim text.