mFA1Murine Fetal Antigen 1
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The MF[alpha]1 sense primer (V190) was labeled with FAM (6-carboxyfluorescein) at the 5' end, the antisense primer (V191) was labeled with tetrachloro-fluorescein (TET) at the 3' end, and Mfa1 sense primers (V290, V1311, V1313) were labeled with 6-carboxy-2', 4', 4', 5', 7', 7'-hexachlorofluorescein (HEX) at the 5' end.
The electrophoretic runs with POP-4 polymer produced a 112-bp DNA fragment for Mfa1 and 97-bp fragment for MF[alpha]1, which were easily distinguished with the GeneScan software by the characteristic peak sizes (Figure 3).
CE-SSCP test under nondenaturing conditions with 3% GeneScan polymer at 30[degrees]C allowed characteristic peak patterns to be detected in MF[alpha]1 and MFa1 genes because of the individual differences within the nueleotide sequences.
Based on our success with MF[alpha]1 sense primer in the detection of characteristic peaks for CnVG, CnVN, and Cg, we labeled Mfa1 sense strands and analyzed MATa strains.