Internal positive control for probe-based nucleic acid molecule assays and methods of making and using thereof

A method of making an internal positive control nucleic acid molecule for a probe-based nucleic acid molecule assay which comprises

• creating a first DNA fragment and a second DNA fragment from a template DNA and first set of primers and a second set of primers;
• creating a third DNA fragment and a fourth DNA fragment from the first DNA fragment and the second DNA fragment with a third set of primers and a second set of primers;
• hybridizing the third DNA fragment and the fourth DNA fragment to obtain a first hybridized DNA;
• using a fifth primer set to create a fifth DNA fragment from the first hybridized DNA;
• using a sixth primer set and a seventh primer set to create a sixth DNA fragment and a seventh DNA fragment from the fifth DNA fragment;
• creating a eighth DNA fragment and a ninth DNA fragment from the sixth DNA fragment and the seventh DNA fragment using a eighth primer set and a ninth primer set;
• hybridizing the eighth DNA fragment and the ninth DNA fragment to obtain a second hybridized DNA;
• creating a tenth DNA fragment and an eleventh DNA fragment from the second hybridized DNA using a tenth set of primers and an eleventh set of primers;
• creating a twelfth DNA fragment and a thirteenth DNA fragment from the tenth DNA fragment and the eleventh DNA fragment using a twelfth set of primers and a thirteenth set of primers;
• hybridizing the twelfth DNA fragment and the thirteenth DNA fragment to obtain the internal positive control nucleic acid molecule.