PRODUCTS AND METHODS FOR TISSUE PRESERVATION
The present application relates to methods for increasing stability of formalin fixed, optionally paraffin embedded biological material. In one example, DNAse inhibitors and/or RNAse inhibitors are combined with the biological material or formalin at the time of fixation or shortly prior.
1 . A composition comprising:
a. a degradation inhibitor, and
b. formalin.
2 . The composition of claim 1 wherein the formalin is phosphate buffered at between 1-37% by mass.
3 . The composition of claim 1 further comprising a solid tissue sample.
4 . (canceled)
5 . The composition of claim 1 wherein the degradation inhibitor comprises a small molecule inhibitor of the phosphoryl transferase superfamily.
6 .- 12 . (canceled)
13 . The composition of claim 1 wherein the degradation inhibitor a reactive oxygen species scavenger.
14 .- 16 . (canceled)
17 . A composition comprising:
a. a small molecule inhibitor of the phosphoryl transferase superfamily, and
b. a reactive oxygen species scavenger.
18 .- 20 . (canceled)
21 . A method for preserving a polynucleotide in a biologic sample comprising:
a. contacting the biologic sample with formalin,
b. contacting the biologic sample with a degradation inhibitor.
22 . The method of claim 21 wherein the contacting the biologic sample with formalin is performed at a temperature between 1° C. and 6° C. or at room temperature.
23 . The method of claim 21 wherein the formalin has a pH in the range of 3 to 10.
24 . The method of claim 21 wherein the degradation inhibitor comprises a small molecule inhibitor of the phosphoryl transferase superfamily.
25 . The method of claim 24 wherein the small molecule inhibitor comprises one or more of (Table 1).
26 . The method of claim 21 wherein the degradation inhibitor comprises a water-soluble reactive oxygen species scavenger.
27 . The method of claim 26 wherein the water-soluble reactive oxygen species scavenger comprises one or more of (table 2).
28 . The method of claim 21 wherein the degradation inhibitor comprises a small molecule inhibitor of the phosphoryl transferase superfamily and a water-soluble reactive oxygen species scavenger.
29 . The method of claim 28 wherein the small molecule inhibitor comprises one or more of (Table 1) and wherein the water-soluble reactive oxygen species scavenger comprises one or more of (table 2).
30 . A method for analyzing a polynucleotide comprising
a. obtaining a biologic sample, wherein the sample has been contacted with formalin solution, contacted with a degradation inhibitor, dehydrated, embedded, and stored;
b. purifying the polynucleotide, and
c. analyzing the polynucleotide.
31 .- 45 . (canceled)
46 . A formalin fixed paraffin embedded tissue containing a plurality of polynucleotides wherein at least 5% of the polynucleotides strands are greater than 300 base pairs.
47 . A polynucleotide recovered from fixed tissue:
a. wherein the polynucleotide strand is at least 300 base pairs,
b. wherein the tissue was fixed in formalin,
c. wherein the formalin comprises a degradation inhibitor.
d. wherein the fixed tissue was embedded in paraffin.
48 .- 53 . (canceled)
54 . A polynucleotide recovered from fixed tissue:
a. wherein the polynucleotide strand is at least 300 base pairs,
b. wherein the tissue was fixed in formalin,
c. wherein the tissue was washed in a ethanol-water wash comprising a water-soluble reactive oxygen species scavenger, and
d. wherein the fixed tissue was embedded in paraffin.
55 . The polynucleotide of claim 54 wherein the reactive oxygen species scavenger comprises one or more of (Table 2).