Compositions and methods for using fixed biological samples
The present disclosure provides compositions and methods for using fixed biological samples in partition-based assays. In at least one embodiment, the disclosure provides a composition comprising a fixed biological sample and an un-fixing agent contained in a partition, such as a discrete droplet. In some embodiments, the disclosure provides un-fixing agent compounds capable of catalytically cleaving crosslinks in fixed biological samples, particularly crosslinked nucleic acids, such as RNA.
1. A composition comprising a compound selected from compound (8), compound (9), compound (10), or a combination thereof:
2. The composition of claim 1 , further comprising a fixed biological sample; optionally, wherein the fixed biological sample is derived from a tissue sample, a biopsy sample, or a blood sample.
3. The composition of claim 2 , wherein the composition is within a partition.
4. The composition of claim 2 , wherein the fixed biological sample comprises a sectioned tissue sample.
5. The composition of claim 1 , further comprising a protease.
6. The composition of claim 5 , wherein the protease is a thermolabile protease.
7. The composition of claim 5 , wherein the protease is a cold-active protease.
8. The composition of claim 1 , wherein the compound is compound (8).
9. The composition of claim 1 , wherein the compound is present as compound (8) formulated at a pH of about 6.5 or about 6.8.
10. A compound having the formula:
11. The compound of claim 10 , wherein the compound is formulated at a pH of about 6.5 or about 6.8.
12. A kit comprising: assay reagents; and an un-fixing agent composition comprising a compound selected from consisting of compound (8), compound (9), compound (10), and a combination thereof:
13. The kit of claim 12 , wherein the compound is compound (8).
14. The kit of claim 12 , wherein the compound is present as compound (8) formulated at a pH of about 6.5 or about 6.8.
15. The composition of claim 1 , wherein the compound is present at a concentration of about 50 mM to about 300 mM.
16. The compound of claim 10 , wherein the compound is formulated at a concentration of about 50 mM to about 300 mM.
17. The kit of claim 12 , wherein the compound is present in the composition at a concentration of about 50 mM to about 300 mM.