Nucleic acid extraction solution and use thereof

Abstract

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 nucleic acid extraction solution comprising a molecule in an amount sufficient to extract nucleic acids from a biological sample, the molecule having the formula R 1O —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. 
     
     
         2 . The solution of  claim 1 , wherein the alkyl group has 1-6 carbon atoms. 
     
     
         3 . The solution of  claim 2 , wherein the alkyl group is selected from the group consisting of methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl, and iso-hexyl. 
     
     
         4 . The solution of  claim 2 , wherein the alkyl group is selected from the group consisting of methyl, ethyl, n-butyl, iso-butyl, sec-butyl, and tert-butyl. 
     
     
         5 . The solution of  claim 1 , wherein R.sub.1 is methyl, ethyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl and R.sub.2 is hydrogen. 
     
     
         6 . The solution of  claim 1 , wherein the molecule is selected from the group consisting of 2-methoxyethanol, 2-ethoxyethanol, and 2-n-butyloxyethanol. 
     
     
         7 . The solution of  claim 1 , wherein the molecule is 2-methoxyethanol. 
     
     
         8 . The solution of  claim 1 , wherein the solution comprises from about 0.5% (v/v) to about 5% (v/v) of the molecule. 
     
     
         9 . The solution of  claim 8 , wherein the solution comprises about 1% (v/v) of the molecule. 
     
     
         10 . The solution of  claim 1 , further comprising a buffering agent. 
     
     
         11 . The solution of  claim 10 , wherein the buffering agent is selected from the group consisting of a Tris buffer, a MOPS buffer, and a borate buffer. 
     
     
         12 . The solution of  claim 1  or  10 , wherein the solution has a pH greater than about 7. 
     
     
         13 . The solution of  claim 12 , wherein the pH is greater than about 7 and less than about 13. 
     
     
         14 . The solution of  claim 1  or  10  further comprising a detergent. 
     
     
         15 . The solution of  claim 14 , wherein the detergent is selected from the group consisting of Tween®, Brij®, and Triton®-X100. 
     
     
         16 . A method of extracting nucleic acid from a biological sample, the method comprising: mixing the sample with a solution comprising a molecule having the formula R 1O —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, so that nucleic acid is released from cells or cellular debris in the sample. 
     
     
         17 . The method of  claim 16 , wherein the alkyl group has 1-6 carbon atoms. 
     
     
         18 . The method of  claim 17 , wherein the alkyl group is selected from the group consisting of methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl, and iso-hexyl. 
     
     
         19 . The method of  claim 17 , wherein the alkyl group is selected from the group consisting of methyl, ethyl, n-butyl, iso-butyl, sec-butyl, and tert-butyl. 
     
     
         20 . The method of  claim 16 , wherein R.sub.1 is methyl, ethyl, n-butyl, iso-butyl, sec-butyl, or tert-butyl and R.sub.2 is hydrogen. 
     
     
         21 . The method of  claim 16 , wherein the molecule is 2-methoxyethanol. 
     
     
         22 . The method of  claim 16 , further comprising a buffering agent. 
     
     
         23 . The method of  claim 22 , wherein the buffering agent is selected from the group consisting of a Tris buffer, a MOPS buffer, and a borate buffer. 
     
     
         24 . The method of  claim 16  or  22 , wherein the solution has a pH greater than about 7. 
     
     
         25 . The method of  claim 24 , wherein the pH is greater than about 7 and less than about 13. 
     
     
         26 . The method of  claim 16  comprising the additional step of heating the mixture to a temperature within the range of from about 50° C. to about 100° C. 
     
     
         27 . The method of  claim 26 , comprising heating the mixture to a temperature of from about 75° C. to about 100° C. 
     
     
         28 . The method of  claim 27 , comprising heating the mixture to a temperature of from about 90° C. to about 100° C. 
     
     
         29 . The method of  claim 16 , wherein the solution comprises about 1% 2-methoxyethanol and borate buffer, pH 9.5. 
     
     
         30 . The method of  claim 16 , comprising the additional step of amplifying a nucleic acid sequence extracted from the sample. 
     
     
         31 . The method of  claim 16 , comprising the additional step of detecting a nucleic acid sequence extracted from the sample. 
     
     
         32 . The method of  claim 30 , wherein the amplification step uses a pair of amplification primers comprising the sequences of SEQ ID NOS: 4 and 5. 
     
     
         33 . The method of  claim 32 , comprising the additional step of detecting the presence of the nucleic acid sequence with a probe comprising the sequence of SEQ ID NO: 3. 
     
     
         34 . The method of  claim 30 , wherein the amplification step uses a pair of amplification primers comprising the sequences of SEQ ID NOS: 8 and 9. 
     
     
         35 . The method of  claim 34 , comprising the additional step of detecting the presence of nucleic acid sequence a probe comprising the sequence of SEQ ID NO: 10. 
     
     
         36 . The method of  claim 16 , wherein the method lacks a chloroform extraction step, a phenol extraction step, a phenol/chloroform extraction step, or an alcohol precipitation step. 
     
     
         37 . The method of  claim 16 , wherein the nucleic acid is a bacterial or viral nucleic acid. 
     
     
         38 . The method of  claim 16 , wherein the biological sample is harvested from a mammal. 
     
     
         39 . The method of  claim 38 , wherein the biological sample comprises cervical cells or cell debris. 
     
     
         40 . The method of  claim 38 , wherein the biological sample comprises breast cells or cell debris.