Method and kit for purifying nucleic acids
Methods for automated extraction of nucleic acids are disclosed. Also disclosed are method and kits for isolating fetal nucleic acids from a plasma sample of a pregnant woman. The method includes flowing the plasma sample through a first filter under conditions that allow binding of the fetal and maternal nucleic acids to the first filter; eluting the fetal and maternal nucleic acids bound to the first filter to produce a concentrated nucleic acid sample; flowing the concentrated nucleic acid sample through a second filter under conditions that allow preferential binding of the maternal nucleic acids to the second filter; and recovering the fetal nucleic acid from the concentrated nucleic acid sample that flow through the second filter.
1. A kit for isolating fetal nucleic acids from maternal nucleic acids in a plasma sample,
comprising:
a pipette tip comprising a self-supporting glass frit filter, wherein the glass frit filter specifically binds to nucleic acids, wherein the glass frit filter has a pore size of 2-220 microns and is not treated or coated with an agent that improves binding of nucleic acid to the glass frit filter,
a first binding buffer formulated to be mixed with a first sample and provide a first binding mixture having about 17-25% v/v of an aliphatic alcohol and a chaotropic salt at a concentration of between about 0.5 M to about 4.0 M; and
a second binding buffer formulated to mixed with a second sample and provide a second binding mixture having about 0-10% v/v of an aliphatic alcohol and a chaotropic salt at a concentration of between about 1 M to about 4.0 M.
2. The kit of claim 1 , comprising:
a first pipette tip comprising a first glass fit filter and having a tip volume of 10-50 ml; and
a second pipette tip comprising a second glass frit filter and having a tip volume of 0.5-2 ml.
3. The kit of claim 2 , wherein the first glass fit filter has a pore size of 16-40 microns and the second glass fit filter has a pore size of 4-5.5 microns.
4. The kit of claim 1 , wherein the glass frit filter comprises a fused glass fit comprising a first section having a first pore size and second section having a second pore size.
5. The kit of claim 4 , wherein the first section has a pore size of 1-160 microns and the second section has a pore size of 16-40 microns, or where in the first section has a pore size of 16-40 microns and the second section has a pore size of 4-5.5 microns.
6. The kit of claim 1 , wherein the first section has a pore size of 100-160- microns and the second section has a pore size of 16-40 microns, or wherein the first section has a pore size of 16-40 microns and the second section has a pore size of 4-5.5 microns.
7. The kit of claim 4 , where in the glass first glass frit filter comprises a fused glass frit comprising a first section having a first pore size and second section having a second pore size.
8. The kit of claim 7 , wherein the first section has a pore size of 100-160- microns and the second section has a pore size of 16-40 microns, or wherein the first section has a pore size of 16-40 microns and the second section has a pore size of 4-10 microns.
9. The kit of claim 1 , wherein the first binding buffer is formulated to be mixed with the first sample and provide the first binding mixture having about 22.5% v/v of an aliphatic alcohol and a chaotropic salt at a concentration of about 1.8M.
10. The kit of claim 1 , wherein the second binding buffer is formulated to be mixed with the second sample and provide the second binding mixture having about 4.7% v/v of an aliphatic alcohol and chaotropic salt at a concentration of about 3.4M.
11. The kit of claim 1 , wherein the aliphatic alcohol in the first binding buffer comprises ethanol or isopropanol.
12. The kit of claim 1 , wherein the aliphatic alcohol in the second binding buffer comprises ethanol or isopropanol.
13. The kit of claim 1 , wherein the a chaotropic salt in the first binding buffer comprises guanidine isothiocyanate or guanidine hydrochloride.
14. The kit of claim 1 , wherein the a chaotropic salt in the second binding buffer comprises guanidine isothiocyanate or guanidine hydrochloride.
15. The kit of claim 2 , further comprising a third pipette tip comprising a third glass frit filter and having a tip volume of 0.2-2 ml.
16. The kit of claim 15 , wherein the third glass frit filter has a pore size of 16-40 microns.
17. The kit of claim 15 , wherein the third glass frit filter a pore size of 4-5.5 microns.
18. The kit of claim 15 , wherein the third glass frit filter has a pore size of 4-10 microns.
19. The kit of claim 15 , wherein the third glass frit filter is a fused filter having a first section with a pore size of 100-160 microns and a second section with a pore size of 16-40 microns.
20. The kit of claim 15 , wherein the third glass frit filter is a fused filter having a first section with a pore size of 16-40 microns and a second section with a pore size of 4-10 microns.