Compositions for in situ nucleic acid analysis
Sample preparation processes for in situ RNA or DNA analysis, methods and compositions therefor are provided. Processes provided herein allow DNA or RNA analysis to be carried out in the same tube or on an aliquot of the prepared sample without centrifugation or extraction. The preparation process can be carried out at room temperature in as little as seven minutes and is amenable to high throughput processing using manual or robotic platforms.
1. A composition comprising: a polypeptide having deoxyribonuclease activity and a surfactant that substantially lacks fluorescence between 300 nm and 750 nm when in use for in situ analysis of nucleic acid, wherein the composition is substantially free of a cation chelator.
2. The composition of claim 1 further comprising a ribonuclease inhibitor.
3. The composition of claim 1 further comprising a reverse transcriptase.
4. The composition of claim 1 further comprising a polymerase.
5. The composition of claim 1 wherein the surfactant comprises octylphenol ethoxylate having an average of 7.5 ethoxylate groups.
6. The composition of claim 1 wherein the surfactant comprises octylphenol ethoxylate having an average of 9.5 ethoxylate groups.
7. The composition of claim 1 wherein the surfactant comprises dodecyl alcohol polyoxyethylene ether having an average of 9.5 ethoxylate groups.
8. The composition of claim 1 wherein the surfactant comprises octylphenolpoly(ethyleneglycolether).
9. The composition of claim 1 comprising less than 0.1 mM cation chelator.
10. The composition of claim 1 comprising less than 0.2 mM cation chelator.
11. The composition of claim 1 comprising less than 0.5 mM cation chelator.
12. The composition of claim 1 comprising less than 1 mM cation chelator.
13. A composition comprising: a polypeptide having deoxyribonuclease activity and a surfactant that substantially lacks fluorescence between 300 nm and 750 nm when in use for in situ analysis of nucleic acid, wherein the composition is substantially free of a cation chelator, wherein the surfactant comprises octylphenol ethoxylate having an average of 7.5 ethoxylate groups, and wherein the composition comprises less than 1 mM cation chelator.
14. The composition of claim 13 comprising less than 0.5 mM cation chelator.
15. The composition of claim 13 comprising less than 0.2 mM cation chelator.
16. The composition of claim 13 comprising less than 0.1 mM cation chelator.
17. A composition comprising: a polypeptide having deoxyribonuclease activity and a surfactant that substantially lacks fluorescence between 300 nm and 750 nm when in use for in situ analysis of nucleic acid, wherein the composition is substantially free of a cation chelator, wherein the surfactant comprises octylphenol ethoxylate having an average of 9.5 ethoxylate groups, and wherein the composition comprises less than 1 mM cation chelator.
18. The composition of claim 17 comprising less than 0.5 mM cation chelator.
19. The composition of claim 17 comprising less than 0.2 mM cation chelator.
20. The composition of claim 17 comprising less than 0.1 mM cation chelator.