TABLE 2.
RNA Yield and PCR Results by Donor.
| Donor | Total RNA Yield (μg/μl) |
260/280 Ratio |
PCR Products | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| PMCA1 | PMCA2 | PMCA3 | PMCA4 | |||||||
| A | B/C | A | B/C* | A | B/C | A | B/C | |||
| 03-029 | 16.47 | 2 | − | + | − | −/− | − | − | + | + |
| 03-030 | 18.9 | 2 | − | + | − | −/− | − | + | + | + |
| 03-031 | 5.86 | 1.98 | − | + | − | −/+ | − | − | + | + |
| 03-055 | 9.9 | 2 | − | + | NE | −/− | − | − | + | + |
| 03-065 | 20 | 1.7 | − | NE | − | −/− | − | NE | + | NE |
Splice variants of PMCAs expressed in hCE were determined by PCR amplification of hCE using PMCA isoform gene-specific primers that flanked splice sites A and B/C. NE indicates a sample was not evaluated secondary to insufficient amount of sample. (−) indicates that no PCR products were detected. (+) indicates PCR products were detected; bands were purified and their identity as PMCAs determined by sequencing.
*
PCR at splice site B/C for PMCA2 was done using two different primer pairs (2cap/2cam and P2pBeWo/2cam). In four donors, no reaction products were detected using either primer set (−/−). PCR using the primer pair P2pBeWo/2cam yielded products of the expected size in Donor 03-031.(−/+).