Methods and compositions for measuring beta cell death
Provided herein are, inter alia, nucleic acids, methods, and kits for detecting unmethylated DNA in body fluid sample of a subject. The disclosure includes compositions, methods, and kits for detecting unmethylation at a CpG site in an insulin gene promoter of a subject.
1. A nucleic acid comprising SEQ ID NO: 1, 2, 3, or 4.
2. A nucleic acid complex comprising the nucleic acid of claim 1 , wherein said SEQ ID NO: 1, 2, 3, or 4 is contacted to a complementary deoxyribonucleic acid (DNA) sequence to form a hybridized nucleic acid complex, wherein said complementary DNA sequence comprises uridine.
3. The nucleic acid complex of claim 2 , wherein said nucleic acid is hybridized to a CpG site of said complementary DNA sequence.
4. A deoxyribonucleic acid (DNA) comprising a uridine that is hybridized to a first nucleic acid and a second nucleic acid each independently comprising SEQ ID NO: 1, 2, 3, or 4, to form a hybridized DNA with SEQ ID NO: 1, 2, 3, or 4, wherein said first and said second nucleic acids do not simultaneously comprise the same SEQ ID NO: 1, 2, 3, or 4.
5. The DNA of claim 4 , wherein said first nucleic acid comprises SEQ ID NO: 1 and said second nucleic acid comprises SEQ ID NO: 2.
6. The DNA of claim 4 , wherein said first nucleic acid comprises SEQ ID NO: 3 and said second nucleic acid comprises SEQ ID NO: 4.
7. The DNA of claim 4 , wherein said DNA is derived from a biological sample.
8. The DNA of claim 7 , wherein said biological sample is blood.
9. The DNA of claim 8 , wherein said blood is whole blood, plasma, or serum.
10. A kit comprising a first nucleic acid and a second nucleic acid each independently comprising SEQ ID NO: 1, 2, 3, 4, 5, or 6, wherein said first and said second nucleic acids do not simultaneously comprise the same SEQ ID NO: 1, 2, 3, 4, 5, or 6.
11. The kit according to claim 10 , wherein said first nucleic acid comprises SEQ ID NO: 1 and said second nucleic acid comprises SEQ ID NO: 2.
12. The kit according to claim 10 , wherein said first nucleic acid comprises SEQ ID NO: 3 and said second nucleic acid comprises SEQ ID NO: 4.
13. The kit according to claim 10 , wherein said first nucleic acid comprises SEQ ID NO: 5 and said second nucleic acid comprises SEQ ID NO: 6.
14. The kit according to claim 10 , further comprising: enzymes, reagents for deamination of cytosine, buffers, vials, plasmid vectors, control DNA, devices for collecting blood and/or tissue samples, or reagents for labeling DNA, or any combinations thereof.
15. The kit according to claim 14 , wherein the enzymes are selected from the group consisting of: thermostable DNA polymerase enzymes, restriction enzymes, and combination thereof.