Assay for human DNA for gender determination
A method for determining gender from a human DNA sample. The loci of Alu element insertion is selected, amplified and evaluated in terms of size of the fragment. The gender assay utilizes AluSTXa for the X chromosome, AluSTYa for the Y chromosome, or both AluSTXa and AluSTYa, to reduce the possibility of error to a negligible quantity. The inserted chromosome yields a large fragment when the homologous region is amplified. The males are distinguished as having two DNA amplicons present, while females have only a single amplicon. The kit adapted for carrying out the method includes a pair of primers to amplify the locus and optionally polymerase chain reaction regents.
1. A primer pair for a sex determination of a human DNA sample, said primer pair selected from the group consisting of the following primer pairs:
a primer pair comprising: a forward primer having the sequence consisting of:
5′-ACTGCAGTCCGCAGTCCGG-3′ (SEQ ID NO: 1); and
a reverse primer having the sequence consisting of:
5′-CCACTGACATATTATCAGCCTTATTAC-3′ (SEQ ID NO: 2);
a primer pair comprising: a forward primer having the sequence consisting of:
5′-TGAAGAAATTCAGTTCATAGCTTGT-3′ (SEQ ID NO: 3); and
a reverse primer having the sequence consisting of:
5′-CAGGAGATCCTGAGATTATGTGG-3′ (SEQ ID NO: 4); and
a primer pair comprising: a forward primer consisting of:
5′-AAACATGGCATGGGCCTCT-3′ (SEQ ID NO: 5); and
a reverse primer consisting of:
5′-TTTTAAAGCCACAGACCTCTCTCT-3′ (SEQ ID NO: 6).
2. The primer pair of claim 1 , wherein each of the primer pair has the sequence selected from the group consisting of:
(SEQ ID NO: 1)
Forward 5′-ACTGCAGTCCGCAGTCCGG-3′;
and
(SEQ ID NO: 2)
Reverse 5′-CCACTGACATATTATCAGCCTTATTAC-3′.
3. The primer pair of claim 1 , wherein each of the primer pair has the sequence selected from the group consisting of:
(SEQ ID NO: 3)
Forward 5′-TGAAGAAATTCAGTTCATAGCTTGT-3′;
and
(SEQ ID NO: 4)
Reverse 5′-CAGGAGATCCTGAGATTATGTGG-3′.
4. The primer pair of claim 1 , wherein each primer of the primer pair has the sequence selected from the group consisting of:
(SEQ ID NO: 5)
Forward 5′-AAACATGGCATGGGCCTCT-3′;
and
(SEQ ID NO: 6)
Reverse 5′-TTTTAAAGCCACAGACCTCTCTCT-3′.
5. A kit for determining gender from a sample, comprising:
a pair of primers capable of amplifying a selected locus of a human DNA in an amplification reaction, said selected locus being in a non-combining X-Y homologous region and selected from the group consisting of a locus in AluSTYa on the Y chromosome and a locus in AluSTXa on the X chromosome, said region containing a monomorphic Alu insertion in one of the X chromosome and the Y chromosome, said pair of primers selected from the group consisting of the following primer pairs:
a primer pair comprising a forward primer consisting of 5′-ACTGCAGTCCGCAGTCCGG-3′ (SEQ ID NO: 1) and a reverse primer having the sequence consisting of 5′-CCACTGACATATTATCAGCCTTATTAC-3′ (SEQ ID NO: 2);
a primer pair comprising a forward primer consisting of 5′-TGAAGAAATTCAGTTCATAGCTTGT-3′ (SEQ ID NO: 3) and a reverse primer consisting of 5′- CAGGAGATCCTGAGATTATGTGG-3′ (SEQ ID NO: 4); and
a primer pair comprising a forward primer having the sequence consisting of 5′-AAACATGGCATGGGCCTCT-3′ (SEQ ID NO: 5) and a reverse primer having the sequence consisting of 5′-TTTTAAAGCCACAGACCTCTCTCT-3′ (SEQ ID NO: 6); and
a polymerase chain reaction reagent comprising a polymerase catalyzing the amplification of the selected locus.
6. The kit of claim 5 , wherein the selected locus is AluSTYa locus on the Y chromosome.
7. The kit of claim 5 , wherein each of the primer pair has the sequence selected from the group consisting of:
(SEQ ID NO: 1)
Forward 5′-ACTGCAGTCCGCAGTCCGG-3′;
and
(SEQ ID NO: 2)
Reverse 5′-CCACTGACATATTATCAGCCTTATTAC-3′.
8. The kit of claim 5 , wherein the selected locus is AluSTXa locus on the X chromosome.
9. The kit of claim 5 , wherein each of the primer pair has the sequence selected from the group consisting of:
(SEQ ID NO: 3)
Forward 5′-TGAAGAAATTCAGTTCATAGCTTGT-3′;
and
(SEQ ID NO: 4)
Reverse 5′-CAGGAGATCCTGAGATTATGTGG-3′.
10. The kit of claim 5 , wherein the selected locus comprises AluSTYa locus and AluSTXa locus.
11. The kit of claim 5 , wherein said pair of primers comprises:
a first primer pair comprising a forward primer consisting of 5′-TGAAGAAATTCAGTTCATAGCTTGT-3′ (SEQ ID NO: 3) and a reverse primer consisting of 5′-CAGGAGATCCTGAGATTATGTGG-3′ (SEQ ID NO: 4); and
a second primer pair comprising a forward primer consisting of 5′-ACTGCAGTCCGCAGTCCGG-3′ (SEQ ID NO: 1) and a reverse primer consisting of 5′-CCACTGACATATTATCAGCCTTATTAC-3′ (SEQ ID NO: 2).
12. A kit for determining gender from a sample, comprising:
a pair of primers comprising a forward primer consisting of 5′-AAACATGGCATGGGCCTCT-3′ (SEQ ID NO: 5) and a reverse primer consisting of 5′-TTTAAAGCCACAGACCTCTCTCT-3′ (SEQ ID NO: 6); and
a polymerase chain reaction reagent comprising a polymerase catalyzing the amplification of the alleles.