Polymer particles, nucleic acid polymer particles and methods of making and using the same
The disclosure relates to methods of making polymer particles, said methods including the steps of: making an aqueous gel reaction mixture; forming an emulsion having dispersed aqueous phase micelles of gel reaction mixture in a continuous phase; adding an initiator oil comprising at least one polymerization initiator to the continuous phase; and performing a polymerization reaction in the micelles. Further, the initiator oil is present in a volume % relative to a volume of the aqueous gel reaction mixture of between about 1 vol % to about 20 vol %. The disclosure also relates to methods of making nucleic acid polymer particles having the same method steps and wherein the aqueous gel reaction mixture includes a nucleic acid fragment, such as a primer.
1. A method of making polymer particles, said method comprising:
forming an emulsion comprising a dispersed aqueous phase of an aqueous reaction mixture in an immiscible continuous phase, wherein the aqueous reaction mixture includes an acrylamide monomer or a derivative thereof, a bis-acrylamide, and a nucleic acid fragment;
adding an initiator oil comprising at least one polymerization initiator to the continuous phase, wherein the initiator oil is a saturated solution of the at least one polymerization initiator; and
performing a polymerization reaction to form polymer particles conjugated to the nucleic acid fragment;
wherein the initiator oil is present in a volume % relative to a volume % of the gel reaction mixture of between about 1 vol % to about 20 vol %.
2. The method of claim 1 , wherein the initiator oil is present in a volume % relative to a volume % of the gel reaction mixture of between about 2 vol % to about 10 vol %.
3. The method of claim 1 , wherein the polymerization initiator is azobisisobutyronitrile (AIBN).
4. The method of claim 1 , wherein the method yields a polymer particle packed density of at least about 5%.
5. The method of claim 1 , wherein the aqueous reaction mixture includes a second nucleic acid fragment.
6. The method of claim 1 , wherein the bis-acrylamide is included in a range of 5% to 20% relative to the total monomer content.
7. The method of claim 1 , wherein the amount of monomer in the aqueous reaction mixture is sufficient to provide a total monomer content in the polymer particles in a range of 3% to 20%.
8. The method of claim 7 , wherein the range is 5% to 10%.
9. The method of claim 1 , wherein the polymer particles have an average particle size of less than 5 micrometers.
10. The method of claim 9 , wherein the polymer particles have an average particle size of less than 1.5 micrometers.
11. The method of claim 9 , wherein the polymer particles have a coefficient of variance of less than 15%.
12. The method of claim 1 , wherein the initiator oil includes a diethylhexyl carbonate.
13. The method of claim 1 , wherein the initiator oil includes mineral oil.
14. The method of claim 1 , wherein performing the polymerization reaction includes heating the emulsion to a temperature of at least 50° C. while maintaining the emulsion.
15. The method of claim 14 , wherein the temperature is at least 75° C.
16. The method of claim 1 , further comprising quenching the polymerization reaction following performing the polymerization reaction following a spike in temperature profile.
17. The method of claim 1 , further comprising quenching the polymerization reaction immediately following the appearance of polymer particles.
18. The method of claim 17 , wherein quenching the polymerization reaction includes cooling the emulsion to a temperature not greater than room temperature.
19. The method of claim 17 , wherein quenching the polymerization reaction includes cooling in an ice bath or a freezer.