Dried compositions containing flap endonuclease
There is disclosed a composition of an aqueous solution comprising, consisting or consisting essentially of a flap endonuclease, a bulking agent and an organic buffer, wherein the aqueous solution has an inorganic salt concentration of 5 mM or less and wherein the composition is substantially free of glycerol.
1. A kit for use in performing a nucleic acid based assay, the kit comprising a first vessel containing adried composition, and a second vessel containing a reconstitution solution; wherein the dried composition comprises a bulking agent, a flap-endonuclease, dNTPs, a chelating agent, 3-(N-morpholino)propanesulfonic acid (MOPS) or tris(hydroxymethyl)aminomethane (Tris), and contains substantially no magnesium ions; and wherein the reconstitution solution comprises MgCl 2 at a concentration from about 3.8 mM to about 4.4 mM.
2. The kit of claim 1 , wherein the first vessel is a multiwell plate, wherein one or more wells of the multiwell plate contain a unit dose of the dried composition.
3. The kit of claim 1 , wherein the percent mass of inorganic salt is 0.35% or less, 0.311% or less, 0.249% or less, 0.186% or less, 0.124% or less, 0.062% or less, or 0.031% or less.
4. The kit of claim 1 , wherein the first and second vessels are configured for use in a device adapted for automated transfer of the reconstitution solution from the second vessel into the first vessel.
5. The kit of claim 1 , wherein the bulking agent is selected from the group consisting of: raffinose, sucrose, mannitol, trehalose, and combinations thereof.
6. The kit of claim 5 , wherein the bulking agent is raffinose, trehalose or a combination of raffinose and trehalose.
7. The kit of claim 1 , wherein the dried composition further comprises one or more oligonucleotides selected from the group consisting of: an amplification oligomer, a detection probe, a signal probe, a FRET probe, and invader probe, and combinations thereof.
8. The kit of claim 1 , wherein the dried composition further comprises at least two probe oligonucleotides capable of annealing to a target nucleic acid to form a three-dimensional structure that can be recognized by the flap endonuclease.
9. The kit of claim 1 , wherein the dried composition further comprises a reverse transcriptase or an RNase inhibitor.
10. The kit of claim 1 , wherein the dNTPs include dTTP and dUTP.
11. The kit of claim 1 , wherein the chelating agent is selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), ethylenediamine-N,N′-disuccinic acid (EDDS), methylglycindiacetic acid (MGDA), diethylene triamine pentaacetic acid (DTPA), and combinations thereof.
12. The kit of claim 1 , wherein the dried composition further contains a reverse transcriptase and an RNase inhibitor.
13. The kit of claim 12 , wherein the percent mass of inorganic salt is 0.35% or less, 0.311% or less, 0.249% or less, 0.186% or less, 0.124% or less, 0.062% or less, or 0.031% or less.
14. The kit of claim 12 , wherein the bulking agent is selected from the group consisting of: raffinose, sucrose, mannitol, trehalose, and combinations thereof.
15. The kit of claim 14 , wherein the bulking agent is raffinose, trehalose, or a combination of raffinose and trehalose.
16. The kit of claim 12 , wherein the dried composition further comprises an oligonucleotide selected from the group consisting of: an amplification oligomer, a detection probe, a signal probe, a FRET probe, and invader probe, and combinations thereof.
17. The kit of claim 12 , wherein the dNTPs include dTTP and dUTP.
18. The kit of claim 12 , wherein the chelating agent is selected from the group consisting of EDTA, EGTA, EDDS, MGDA, DTPA, and combinations thereof.
19. The kit of claim 12 , wherein the first vessel is a multiwell plate comprising, wherein one or more wells of the multiwell plate contain a unit dose of the dried composition.
20. The kit of claim 12 , wherein the first and second vessels are configured for use in a device adapted for automated transfer of the reconstitution solution from the second vessel into the first vessel.
21. A kit for use in performing a non-nucleic acid based assay, the kit comprising a first vessel containing a dried composition and a second vessel containing a reconstitution solution; wherein the dried composition comprises a bulking agent, a flap-endonuclease, a DNA polymerase, dNTPs, a chelating agent, 3-(N-morpholino)propanesulfonic acid (MOPS) or tris(hydroxymethyl)aminomethane (Tris), and contains substantially no magnesium ions; and wherein the reconstitution solution comprises MgCl 2 at a concentration from about 3.8 mM to about 4.4 mM.