SE33 MUTATIONS IMPACTING GENOTYPE CONCORDANCE
Disclosed are primer set compositions, methods and kits for human identification using the highly complex sequence locus, SE33 (ACTBP2) in single and multiplex PCR reactions. Additionally, disclosed are three newly discovered single nucleotide polymorphisms (SNPs) within the SE33 locus that can cause discordance seen as mobility shift or allelic dropout. Also disclosed are kits useful in human identification.
1 . An amplification primer set for SE33 locus comprising:
a. a first primer capable of annealing to a primer-binding site 5′ of SE33's STR region; and
b. a second primer, a reverse primer, capable of annealing to a first primer-binding site 3′ of the SE33 STR region, wherein the first primer-binding site comprises one variant SNP, wherein the variant SNP is selected from one of nucleotide positions 316, 317 and 324 as illustrated in SEQ ID NO:3.
2 . The amplification primer set of claim 14 , further comprising at least a third primer, a second reverse primer, capable of annealing to a second primer-binding site 3′ of the SE33 STR region.
3 . The amplification primer set of claim 15 , wherein the first reverse primer binding site and the second reverse primer binding sites 3′ of the SE33 STR region share at least 10% sequence identity.
4 . The amplification primer set of claim 15 , wherein the first reverse primer binding site and the second reverse primer binding site 3′ of the SE33 STR region are different.
5 . The amplification primer set of claim 15 , wherein the 3′ terminus of the third primer extends at least 1 to at least 30 nucleotides towards the 5′ end of a template from the variant SNP position.
6 . The amplification primer set of claim 15 , wherein at least one reverse primer can further comprise at least one nucleotide analog or at least one universal base or at least one modified nucleotide or at least one deaza-G nucleotide or can be a degenerate reverse primer.