Based on the findings, that there are specific REnzyme cutting sites present or absent defining membership to a specific clade ( see "Genetic testing - the idea")
subsequent PCR-reactions for the most abundant clades were performed.
For the Clade U we amplified the mitochondrial DNA from
pos. 12104 to 12338, yielding a PCR-product of 240 nucs.
Protocolls see here!
Part of the mitochondrial DNA (pos. 12104 to 12338) amplified by PCR:
Left lane: 50-bp-Ladder, middle and right lane are successful PCR samples right
lane carries excess primers, PCR-Product about 240 nucs.
There are two Variants in Nucleotide Sequence accountig for Clade-U-membership:
Variant A: with Restriction-Site "gantg" of REnzyme HinfI

Variant B: without Restriction-Site

All members of the Clade U, i.e. all descendants of Ursula, belong to Variant A.
So all we have to do, is trying to cut the PCR-Products by the REnzme HinfI.

Ursula (incomplete restriction shows position of uncut PCR-product)

