TABLE 2.
Primer pairs used to genotype Lrp1 knock-in ES cell lines and mice
| Primer pair | Primersa | Amplimer size (bp) | PCR result
|
|
|---|---|---|---|---|
| Before RMCE | After RMCE | |||
| AB | CACAGTGTCCCCTCCCCTCTC | 453 | + | − |
| GCCCTGCCATAGCCACTG | ||||
| CD | TTAATATGCGAAGTGGACCTC | 449 | + | − |
| GAGTCATAAGGAGTCTGGAG | ||||
| EF | GTTCCCTCCATGCCCTGACA | 487 (wt) | + | + |
| GGAGCCTGCCGGAGTGAGA | 577 | − | + | |
| GH | CTGAAGGCTCTTTACTATTGC | 405 | − | + |
| GAGTCATAAGGAGTCTGGAG | ||||
| IJb | ACTTCCAACGCCCACGCGGC | 776 | ||
| GGGGCATCTGGAGGGGGTGTTG | ||||
| KLc | CCGCTGATGGCTCCCGACAAT | 466 | ||
| TACATCTTGGCGGTAGCGGC | ||||
| MNd | GCCTACCAACTTCACCGCCG | 442 | ||
| GCCGCCAGCTTCCAGGGGTATG | ||||
| OPe | AATCCTGCTCCTCTCCCTCTC | 398 | ||
| TATCTCCGCAGTCGTTGTCGT | ||||
a
Primer pairs IJ to MN were used in mutation-specific PCRs. Nucleotides in bold represent mutated residues not present in wild-type Lrp1. wt, wild type.
b
PCR result was positive only in the case of a furin site mutation.
c
PCR result was positive only in the case of an NPXY1 mutation.
d
PCR result was positive only in the case of an NPXY2 mutation.
e
Genomic probe A for Southern blot analysis.