Table 5. Sequence analysis of oligonucleotide donors.
Oligonucleotide | 30 bp Upstream | LoxP Sequence | 30 bp Downstream |
---|---|---|---|
LoxP4 oligonucleotide | TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAA__TCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACAT_AGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTT_GGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | _TAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGT_AT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
TCCATTACATTAGCATGTGCAGTGGCCATG | ATAACTTCGTATAGCATACATTATACGAAGTTAT | GTCAGGGTGTGGAGCGTTTTGGGAGCTTTA | |
LoxP5 oligonucleotide | TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAG_ATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGAC_TGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCATCTAACAGCCATCTCTACCTCC | |
TTGCAGAGGACCTGGGTTTGGTTCCTAGCT | ATAACTTCGTATAGCATACATTATACGAAGTTAT | CACTGGCACCTAACAGCCATCTCTACCTCC |
Both loxP4 (194 bp, top) and loxP5 (193 bp, bottom) oligonucleotide donors were cloned, and 10 clones of loxP4 oligo donor and nine clones of loxP5 donor were sequenced. A series of 1- or 2-bp deletions were identified in both oligonucleotides with a 1-bp substitution was present in the loxp5 oligonucleotide. Observed mutation or substitutions are shown in gray.