Confirmation of Predicted Splice Variants Using RT-PCR
| A | ||
| Accession | Primer name | Primer sequence |
| D13906 | D13906_1S | TCGGTGGAGCTCCACTCTCCT |
| D13906 | D13906_2A | GAAGGAGTCTATGCACTTCTCC |
| D13906 | D13906_3A* | GCTCAGGATCATGCAAACTCTG |
| J03588 | J03588_1S | TCTGAAGAGACCTGGCACACTC |
| J03588 | J03588_2A* | GTGTGATCATCTGAGGGAAGGC |
| J03588 | J03588_3A | AGCCACACTCCAGCTACAAAGG |
| M32801 | M32801_1S* | CATCGGACCTGCCTATGCCATC |
| M32801 | M32801_2S | GTGTGGAGAAGCTGGGAATTCC |
| M32801 | M32801_3A | TCCCACGAAGATGGCTCCTGTA |
| M34007 | M34007_1S | TGATCAGATGGACAACGCCAAG |
| M34007 | M34007_2A* | CACTTCTCAGTGGGTCTTGGAT |
| M34007 | M34007_3A | TTCTGGGTGTATAGTGTATGGTG |
| X07467 | X07467_1S | CTTTGGGACCATAGGCCTTAGC |
| X07467 | X07467_2A | CTCAGGGAAGTGTGGTTTGGTC |
| X07467 | X07467_3A* | TAAGGCTAGTGTGGCTATGGGC |
| D30035 | D30035_1S | GGATTCTCACTTCTGTCATCTGGC |
| D30035 | D30035_2S* | CAAGCGCACCATTGCTCAGGAT |
| D30035 | D30035_3A | TTTCTTCTGGCTGCTCAAAGCTG |
| B | |||||||
| Accession | Primer pair | Tissue 1 | Tissue 2 | ||||
| name | predicted (bp) | PCR (bp) | name | predicted (bp) | PCR (bp) | ||
| D13906 | D13906_1S,D13906_2A | Kidney | 350 | ∼350 | Placenta | 350 | ∼350 |
| D13906 | D13906_1S,D13906_3A | Kidney | 450 | ∼450 | Placenta | no signal | ∼450 |
| J03588 | J03588_1S,J03588_2A | Liver | 274 | ∼270 | Kidney | no signal | ∼270 |
| J03588 | J03588_1S,J03588_3A | Liver | 360 | ∼380 | Kidney | <380 | ∼380 |
| M32801 | M32801_1S,M32801_3A | Liver | 401 | ∼400 | Kidney | no signal | no signal |
| M32801 | M32801_2S,M32801_3A | Liver | 291 | ∼290 | Kidney | 291 | ∼290 |
| M34007 | M34007_1S,M34007_2A | Testis | no signal | no signal | Bladder | 424 | ∼425 |
| M34007 | M34007_1S,M34007_3A | Testis | <542 | ∼420 | Bladder | 542 | ∼550 |
| X07467 | X07467_1S, X07467_2A | Sm Intestine | 330 | ∼330 | Liver | 330 | ∼330 |
| X07467 | X07467_1S, X07467_3A | Sm Intestine | 410 | ∼400 | Liver | no signal | no signal |
| D30035 | D30035_1S,D30335_3A | Kidney | 410 | ∼400 | Heart | <410 | ∼400 |
| D30035 | D30035_2S, D30035_3A | Kidney | 324 | no signal | Heart | no signal | weak signal |
-
(A) The list of PCR primers and their corresponding gene (Accession). An asterisk indicates that the primer is located within the predicted alternative splicing region. All primer sequences are from 5′ to 3′. (B) Comparison of predicted and actual RT-PCR results. Two pairs of primers were used for each gene. For each gene, a pair of different tissue samples was chosen to perform the RT-PCR experiment. The Predicted column indicates the predicted size of PCR product (or no signal) based on the Splice Variant Prediction Algorithms. The PCR column shows the size of the actual RT-PCR product (or no signal) revealed by the experiment. The genes with correlated result between predicted and actual PCR are highlighted in bold.
