Nucleic acid extraction solution and use thereof
View Patent ↗Disclosed are methods and compositions for extracting nucleic acids from a biological sample. In particular, disclosed is a nucleic acid extraction solution together with methods using such a solution for extracting nucleic acid sequences from biological samples containing cells, cellular debris or both. The nucleic acid extraction solution contains a molecule having the formula R 1 O—CH 2 —CH 2 —OR 2 , wherein R 1 and R 2 independently are selected from the group consisting of hydrogen and an alkyl group.
1. A method of extracting nucleic acid from a biological sample, the method comprising: mixing the sample with a solution comprising about 1% 2-methoxyethanol and borate buffer, so that nucleic acid is released from cells or cellular debris in the sample.
2. The method of claim 1 , wherein the solution has a pH greater than about 7.
3. The method of claim 2 , wherein the pH is greater than about 7 and less than about 13.
4. The method of claim 1 comprising the additional step of heating the mixture to a temperature within the range of from about 50° C. to about 100° C.
5. The method of claim 4 , comprising heating the mixture to a temperature of from about 75° C. to about 100° C.
6. The method claim 5 , comprising heating the mixture to a temperature of from about 90° C. to about 100° C.
7. The method of claim 1 , comprising the additional step of amplifying a nucleic acid sequence extracted from the sample.
8. The method of claim 1 , comprising the additional step of detecting a nucleic acid sequence extracted from the sample.
9. The method of claim 7 , wherein the amplification step uses a pair of amplification primers comprising the sequences of SEQ ID NOS: 4 and 5.
10. The method of claim 9 , comprising the additional step of detecting the presence of the nucleic acid sequence with a probe comprising the sequence of SEQ ID NO: 3.
11. The method of claim 7 , wherein the amplification step uses a pair of amplification primers comprising the sequences of SEQ ID NOS: 8 and 9.
12. The method of claim 11 , comprising the additional step of detecting the presence of nucleic acid sequence a probe comprising the sequence of SEQ ID NO: 10.
13. The method of claim 1 , wherein the method lacks a chloroform extraction step, a phenol extraction step, a phenol/chloroform extraction step, or an alcohol precipitation step.
14. The method of claim 1 , wherein the nucleic acid is a bacterial or viral nucleic acid.
15. The method of claim 1 , wherein the biological sample is harvested from a mammal.
16. The method of claim 15 , wherein the biological sample comprises cervical cells or cell debris.
17. The method of claim 15 , wherein the biological sample comprises breast cells or cell debris.