A High Resolution HLA Sequence- Specific Primer PCR Reaction Was

Multiple Choice

A high resolution HLA sequence- specific primer PCR reaction was done in a 96 well microtiter plate. Each well contains primers to a different HLA allele and the same primers to a sequence of DNA to be used as an internal control. A negative control consists of distilled water left in an open tube during the specimen processing. After PCR, the contents of each well were electrophoresed and visualized on gel. Which finding automatically indicates a potentially invalid reaction that requires repeat analysis?

A) In the well that an HLA- A gene amplified, the control sequence also amplified (i.e., two bands were visible) .
B) Two different alleles for the HLA- A locus were amplified.
C) An allele determined by PCR does not match the allele determined by classic serology.
D) No bands were visualized in the negative control.
E) An allele identified by the high resolution procedure does not match the allele identified by the low- resolution procedure.