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US 2021/0102245 A1Published application
Direct nucleic acid analysis of environmental and biological samples
AssigneeSpartan Bioscience Inc.
InventorsChris Harder, Christine Dobson, Paul Lem, Alan Mears, Jeffrey Do, Ali Khatib
Life of this patent
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Abstract
Methods and apparatuses are described for nucleic acid analysis of environmental water samples and biological samples without the need for purification.
Claims40
A method comprising steps of:
- obtaining an environmental sample comprising a microorganism, wherein the microorganism comprises a nucleic acid;
- concentrating the environmental sample to produce a concentrated sample, wherein the microorganism is concentrated about 2-fold to about 125-fold in the concentrated sample as compared to the environmental sample;
- contacting the concentrated sample with a nucleic acid amplification reagent in a reaction vessel, wherein the concentrated sample is directly contacted with the nucleic acid amplification reagent without any intervening steps;
- and
- performing a nucleic acid amplification reaction on the nucleic acid from the microorganism in the concentrated sample.
2 more elements
A method comprising steps of:
- obtaining a sample comprising a nucleic acid;
- contacting the sample with a nucleic acid amplification reagent in a reaction vessel,
- wherein the sample is directly contacted with the nucleic acid amplification reagent without any intervening steps and
- wherein the nucleic acid amplification reagent comprises a DNA polymerase at a concentration ranging from at least 6 U/reaction to about 42 U/reaction, a primer at a concentration ranging from at least 2.0 μM to about 14 μM and a probe at a concentration ranging from at least 1.9 μM to about 14 μM;
- and
- performing a nucleic acid amplification reaction on the nucleic acid from the sample.
3 more elements
A method comprising steps of:
- obtaining an environmental sample from a source, wherein the environmental sample comprises a microorganism and the microorganism comprises a nucleic acid;
- optionally concentrating the environmental sample to produce a concentrated sample, wherein the microorganism is concentrated about 2-fold to about 125-fold in the concentrated sample as compared to the environmental sample;
- contacting the environmental sample or concentrated sample with a nucleic acid amplification reagent in a reaction vessel, wherein the environmental sample or concentrated sample is directly contacted with the nucleic acid amplification reagent without any intervening steps;
- and
- performing a nucleic acid amplification reaction on the nucleic acid from the microorganism in the environmental sample or concentrated sample, wherein the nucleic acid amplification reaction is completed within less than 1 day from when the environmental sample was originally collected from the source.
2 more elements
A method comprising steps of:
- (a) obtaining a first environmental sample from a source, wherein the environmental sample comprises a microorganism and the microorganism comprises a nucleic acid;
- (b) optionally concentrating the environmental sample to produce a concentrated sample, wherein the microorganism is concentrated about 2-fold to about 125-fold in the concentrated sample as compared to the environmental sample;
- (c) contacting the environmental sample or concentrated sample with a nucleic acid amplification reagent in a reaction vessel, wherein the environmental sample or concentrated sample is directly contacted with the nucleic acid amplification reagent without any intervening steps;
- (d) performing a nucleic acid amplification reaction on the nucleic acid from the microorganism in the environmental sample or concentrated sample, wherein the nucleic acid amplification reaction is optionally completed within less than 1 day from when the environmental sample was originally collected from the source;
- and
- repeating steps (a), (c), (d) and optionally (b) on a second environmental sample from the same source within less than one month.
3 more elements
Dependencies are inferred from the claim text.