Methylated control DNA
Provided herein is technology relating compositions and methods for analysis of methylated DNA from a subject. The technology also relates to use of endogenous methylated DNAs as internal controls for marker gene methylation assays.
1. A composition, comprising:
a) a bisulfite-converted B3GALT6 DNA comprising SEQ ID NO:2 or the full complement of SEQ ID NO:2, and
b) a pair of primers complementary to primer binding sites within SEQ ID NO:2 or the complement thereof.
2. The composition of claim 1 , further comprising a detection probe oligonucleotide, wherein the detection probe oligonucleotide comprises a region that is complementary to a portion of said DNA.
3. The composition of claim 1 , further comprising one or more of:
a detection probe oligonucleotide;
a FRET cassette;
a FEN-1 endonuclease;
a thermostable DNA polymerase.
4. The composition of claim 2 , wherein said detection probe oligonucleotide comprises a reporter molecule.
5. The composition of claim 2 , wherein said detection probe oligonucleotide comprises a flap sequence.
6. The composition of claim 4 , where said reporter molecule comprises a fluorophore.
7. A method, the method comprising:
a) treating DNA from a sample with a bisulfite reagent to produce a bisulfite-converted B3GALT6 DNA;
b) amplifying a region of said bisulfite-converted DNA using a pair of nucleic acid primers; and
c) detecting an amplified B3GALT6 DNA comprising SEQ ID NO:2 or the full complement thereof.
8. The method of claim 7 , further comprising a step of detecting the amplified bisulfite-converted B3ALT6 DNA comprising SEQ ID NO: 2 with a detection probe oligonucleotide.
9. The method of claim 8 , wherein said detection probe oligonucleotide comprises a reporter molecule.
10. The method of claim 8 , wherein said detection probe oligonucleotide comprises a flap sequence.