Method for nucleotide detection
A method of inhibiting light-induced degradation of nucleic acids includes irradiating a portion of the nucleic acids in the presence of a detection solution comprising a polyphenolic compound. A method of detecting a nucleic acid having a fluorescent tag includes irradiating at least a portion of the nucleic acid with light of a suitable wavelength to induce a fluorescence emission and detecting the fluorescence emission. Optionally, the polyphenolic compound is gallic acid, a lower alkyl ester thereof, or mixtures thereof. A kit includes one or more nucleotides, an enzyme capable of catalyzing incorporation of the nucleotides into a nucleic acid strand and a polyphenolic compound suitable for preparing a detection solution.
1. A method of detecting a nucleic acid having a fluorescent tag comprising:
a) irradiating at least a portion of said nucleic acid with light, wherein said light comprises a suitable wavelength to induce a fluorescence emission;
b) detecting said fluorescence emission; and
c) repeating steps a) and b);
wherein said irradiating step is carried out in the presence of a detection solution comprising gallic acid, a lower alkyl ester thereof, or mixtures thereof, and further comprising urea and ascorbic acid or a salt thereof, said detection solution inhibiting light-induced degradation of said nucleic acid.
2. The method of claim 1 , wherein said nucleic acid is in an array of nucleic acids attached to a support.
3. The method of claim 1 , further comprising adding a fluorescently tagged nucleotide to said nucleic acid.
4. The method of claim 1 , further comprising replacing a solution with said detection solution prior to the irradiation step.
5. The method of claim 2 , wherein inhibiting light-induced degradation to said nucleic acid comprises reducing the cleavage of a nucleic acid member from said array.
6. The method of claim 1 , wherein said gallic acid, said lower alkyl ester thereof, or said mixtures thereof is present in a concentration ranging from between about 10 mM to about 200 mM.
7. The method of claim 1 , further comprising one or more compound(s), wherein said one or more compound(s) further enhances preservation of the integrity of said nucleic acid.
8. The method of claim 7 , wherein said one or more compound(s) is isoascorbic acid or a salt thereof.
9. The method of claim 1 , comprising at least 50 cycles repeating step c.
10. The method of claim 1 , comprising repeating said adding and detection steps for a number of cycles in a range from between about 100 cycles to about 1,000 cycles.
11. The method of claim 1 , wherein the presence of said detection solution reduces a detection error rate by greater than 20% relative to a control.
12. The method of claim 11 , wherein isoascorbic acid or a salt thereof further reduce(s) said detection error rate.
13. The method of claim 1 , wherein the presence of said detection solution reduces a detection error rate by greater than 40% relative to a control.
14. The method of claim 13 , wherein isoascorbic acid or a salt thereof further reduce(s) said detection error rate.
15. The method of claim 1 , wherein the presence of said detection solution reduces a detection error rate by greater than 50% relative to a control.
16. The method of claim 15 , wherein isoascorbic acid or a salt thereof further reduce(s) said detection error rate.
17. The method of claim 1 , wherein said irradiation step is conducted in a range from about 360 nm to about 700 nm.
18. The method of claim 1 , wherein said irradiation step is conducted with a light source having power in a range between about 5 to about 500 milliwatts.
19. The method of claim 1 , wherein said irradiation step is conducted for a time period of about 0.1 seconds to about 10 minutes.
20. The method of claim 3 , wherein adding an additional fluorescently tagged nucleotide comprises using a polymerase to add a single nucleotide.