Table 1.
Primers used for genotyping.
| Primer | Sequence | PCR product (bp) | DLST+/+ | DLST+/- |
|---|---|---|---|---|
| primer 5 | 5’-TAGGTTCCTAGGTAGGGATACAGC-3’ | 186 | + | + |
| primer 3 | 5’-CTACTCTCTAACCTACCAAGCTGG-3’ | |||
|
| ||||
| primer 5 | 5’-TAGGTTCCTAGGTAGGGATACAGC-3’ | 210 | - | + |
| LTR-rev | 5’-ATAAACCCTCTTGCAGTTGCATC-3’ | |||
Note: Two sets of primers were used for PCR to genotype the DLST+/+ and DLST+/- mice. Genomic DNA from each pup will be subjected to two PCR. PCR using primer 5/primer 3 will only amplify the wild-type gene and generate product of 186 bp. PCR with primer 5/LTR-rev will only amplify the mutated gene and produce a fragment of 210 bp. Thus, DLST+/+ mice will only give rise to one PCR product of 186 bp. While, DLST+/- mice will have both of 186 and 210 bp fragments from PCR.