Electroelution of oligonucleotides from gel matrices
View Patent ↗The invention can be used to purify and extract a target oligonucleotide from a gel substrate. The current invention extracts the target oligonucleotide so quickly that burn-off against the positive electrode is greatly reduced, yielding high optical density and purity. The need for an osmotic membrane or heavy salt/light salt barrier to capture the oligonucleotide is eliminated. The invention does not require a high salt concentration and therefore does not require a desalting column that is time-consuming and reduces yield.
1. An apparatus for the recovery of a target oligonucleotide in a gel sample, said apparatus comprising:
a) a chamber for placement of the gel sample and a buffer solution;
b) a source of electric current to provide a negative electric current to the chamber;
c) a recovery well separate from the chamber wherein the gel sample forms a bottom of the recovery well, said recovery well capable of holding a buffer solution; and
d) a second source of electric current to provide a positive electric current to the recovery well.
2. An apparatus of claim 1 wherein the apparatus contains more than one chamber and more than one recovery well.
3. An apparatus of claim 1 further comprising a recovery well aspirator.
4. An automated system for the loading and elution of a gel, comprising:
a) a component for the loading of a sample into a gel;
b) a component for the transfer of the gel to the apparatus of claim 1 ; and
c) an aspirator.
5. The automated system of claim 4 wherein the apparatus of claim 1 has more than one chamber and more than one recovery well.
6. A method of removing a nucleic acid sample from a gel, the method comprising:
a) excising a gel portion from the gel, said gel portion containing the nucleic acid sample;
b) providing a negative charge to the gel portion;
c) providing a recovery well, said recovery well being formed by the placement of a surface with an opening, wherein said opening sets against the gel portion and forms a seal to allow a buffer solution to sit in the opening against the gel portion;
d) providing a buffer solution to the opening;
e) providing a positive charge to the buffer solution; and
f) collecting the buffer solution.
7. The method of claim 6 wherein the negative charge and the positive charge are provided for about 90 seconds.
8. The method of claim 6 wherein a current provided by the positive and negative charges is greater than 100 volts.