Table 2. Primers used for Sanger sequencing, genotyping, and qPCR.
| Primer name | Sequence (5’–3’) | GenBank accession number | Nucleotide position |
|---|---|---|---|
| P-WT-F | CCCATAAGTTGTTCCTTGATTTCC | NC_051356.1 | 88350–88373 |
| P-WT-R | CACTGCCTGAATAAGACCCACTC | NC_051356.1 | 88777–88799 |
| P-WT-F’ | CCTTGTGGAAGTAAAACCGAA | NC_051356.1 | 93529–93549 |
| P-WT-R’ | TACAAAGGCCAAGTTCAG | NC_051356.1 | 84492–84510 |
| P-KO-F | CTTCATTCCCTTTGCACGTT | NC_051356.1 | 84092–84111 |
| P-KO-R | TACAAAGGCCAAGTTCAG | NC_051356.1 | 94242–94262 |
| PLS3-KO-F | AAATTCTCCTTGGTTGGCATT | NM_031084.1 | 1507–1527 |
| PLS3-KO-R | TCCAGCTTCACTCAATGTTCC | NM_031084.1 | 1672–1692 |
| PLS3-F | GAAAATGATCCCGATTGCAG | NM_031084.1 | 496–515 |
| PLS3-R | CTCTCATCAATTGTATCGGGAA | NM_031084.1 | 605–626 |
| COL1A1-F | TCCTGACGCATGGCCAAGAA | NM_053304.1 | 164–183 |
| COL1A1-R | CATAGCACGCCATCGCACAC | NM_053304.1 | 289–308 |
| GAPDH-F | TTCAACGGCACAGTCAAGG | NM_017008.4 | 235–253 |
| GAPDH-R | CTCAGCACCAGCATCACC | NM_017008.4 | 331–348 |
Primers PLS3-KO-F/PLS3-KO-R were selected to amplify a fragment spanning from exon 13 to exon 14 of the rat PLS3 cDNA. The qPCR results were used to indicate the expression level of PLS3 E10-16 (knockout region). Primers PLS3-F/PLS3-R were selected to amplify a fragment spanning from exon 5 to exon 6 of the rat PLS3 cDNA. The qPCR results were used to indicate the expression level of PLS3 E1-9 (uneditable region).
F: forward, R: reverse, WT: wild-type, KO: knockout.