NON-CODING RNA FOR DETECTION OF CANCER
The present disclosure relates generally to detection on non-coding RNAS molecules in a sample or diagnosis of subject based upon detection or quantification of non-coding nucleic acid sequences in a sample, specifically to identify and use of molecular biomarkers for cancer including breast cancer.
1 . A method for sequencing a ribonucleic acid (RNA) molecule from a cell-free sample, comprising:
(a) providing said cell-free sample comprising said RNA molecule, wherein said RNA molecule comprises a non-coding sequence;
(b) subjecting said RNA molecule to reverse transcription to generate a complementary deoxyribonucleic acid (cDNA) molecule, wherein said cDNA molecule comprises a sequence corresponding to said non-coding sequence of said RNA molecule; and
(c) subjecting said cDNA molecule or derivative thereof to sequencing-by-synthesis to identify said non-coding sequence.
2 . The method of claim 1 , wherein said RNA molecule is an orphan non-coding RNA (oncRNA).
3 . The method of claim 1 , wherein said RNA molecule comprises T3p or functional fragment thereof.
4 . The method of claim 1 , further comprising, after (a), isolating said RNA molecule from other components of said cell-free sample.
5 . The method of claim 4 , wherein said isolating comprises filtration.
6 . The method of claim 1 , further comprising using a result of said sequencing-by-synthesis to determine an amount of said non-coding sequence in said cell-free sample.
7 . The method of claim 1 , wherein said cell-free sample comprises serum.
8 . The method of claim 1 , wherein said cell-free sample comprises whole blood.
9 . The method of claim 1 , wherein said cell-free sample comprises plasma.
10 . The method of claim 1 , wherein said cell-free sample comprises urine.
11 . The method of claim 1 , wherein said cell-free sample comprises lymph.
12 . The method of claim 1 , wherein said cell-free sample comprises saliva.
13 . The method of claim 1 , wherein the volume of cell-free sample is about 20 microliters to about 2 milliliters.
14 . The method of claim 11 , wherein the volume of cell-free sample is about 100 microliters to about 500 microliters.
15 . The method of claim 1 , wherein said sequencing-by-synthesis generates sequencing reads, which sequencing reads are processed to identify said non-coding sequence.
16 . The method of claim 1 , wherein said RNA molecule has a length of less than 200 nucleotides.
17 . The method of claim 13 , wherein said RNA molecule has a length between 50 and 100 nucleotides.
18 . The method of claim 1 , wherein said non-coding sequence is indicative of cancer.
19 . The method of claim 1 , wherein said cancer is breast cancer.
20 . The method claim 1 , further comprising, after (b), amplifying said cDNA molecule.