Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI10111
Trapped Gene
AC166359.3 (ENSMUSG00000047146)
Vector Insertion
Chr 10: 62275465 - 62276161
Public Clones RRA018 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 62% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000666239 (Chr10:62275466..62276174 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATCTACGCCGATCCTTCCTT Chr10:62275650..62275669 60.06 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000666239 (Chr10:62275466..62276174 -)
Downstram Exon
ENSMUSE00000351741 (Chr10:62275115..62276160 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATCTACGCCGATCCTTCCTT Chr10:62275650..62275669 60.06 50 TCGGCGTAGATCTCCTCACT Chr10:62275637..62275656 59.97 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000666241 Chr10:62300924..62300993 GCTGGATTGAAGGAACAGGA Chr10:62300944..62300963 60.19 50
upstream ENSMUSE00000643009 Chr10:62296286..62296376 GTCAGGGAGCTCATGGAGAC Chr10:62296289..62296308 59.8 60
upstream ENSMUSE00000238699 Chr10:62295642..62295735 GATAGTGTTCACGGGGAAGG Chr10:62295680..62295699 59.4 55
upstream ENSMUSE00000643008 Chr10:62285297..62285508 GTTCGTGAGCGTGTAGACCA Chr10:62285447..62285466 59.9 55
upstream ENSMUSE00000643006 Chr10:62282221..62282371 GAGGAGTGAAAACCCCAGAA Chr10:62282253..62282272 59.11 50
upstream ENSMUSE00000666240 Chr10:62281687..62281782 ATCATTCCAGACCGCAAGAC Chr10:62281703..62281722 60.08 50
upstream ENSMUSE00000643005 Chr10:62279132..62279221 TTACAAGCAGATGGCTCCAG Chr10:62279149..62279168 59.02 50
upstream ENSMUSE00000643003 Chr10:62277184..62277321 TGTCACCTGTTGCATGGATT Chr10:62277237..62277256 59.97 45
upstream ENSMUSE00000643002 Chr10:62276349..62276572 ATTGTCCCGAGGATGAACAA Chr10:62276516..62276535 60.32 45
upstream ENSMUSE00000612637 Chr10:62276326..62276346 No primer for this exon
upstream ENSMUSE00000666239 Chr10:62275466..62276174 ATCTACGCCGATCCTTCCTT Chr10:62275650..62275669 60.06 50

*** Putative Vector Insertion (Chr 10: 62275465 - 62276161) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000351741 Chr10:62275115..62276160 TCGGCGTAGATCTCCTCACT Chr10:62275637..62275656 59.97 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GAGCTTTTCATCAGCCTCATC Chr10:62276122..62276143 59.02 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCACCTCGTGACTGGGAAAA Chr10:62276096..62276116 61.64 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000047146