Genetics Testing by AFLP  for Clade Ursula

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 aPCR-product of 240 nucs.

 

Protocolls see here!

clade_ursula_clip100 

 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 u_12104_12338_12308hinfi

Variant B: without Restriction-Site  

u_12104_12338_12308_hinfi2

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.

Protocolls see here!

re_ursula

Restriction Analysis of the PCR-product of mtDNA (pos. 12104 to 12338): 
Left lane: 50-bp-Ladder, middle/right lane: restriction = Variant A = Member of Clade
Ursula (incomplete restriction shows position of uncut PCR-product)


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