Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31582
Trapped Gene
Ceacam2 (ENSMUSG00000054385)
Vector Insertion
Chr 7: 26315489 - 26323953
Public Clones IST10709C3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 52% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000461821 (Chr7:26323593..26323952 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAGGGAGTCTCGGCCTTTAG Chr7:26323820..26323839 59.84 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000461821 (Chr7:26323593..26323952 -)
Downstram Exon
ENSMUSE00000460573 (Chr7:26315490..26315774 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAGGGAGTCTCGGCCTTTAG Chr7:26323820..26323839 59.84 55 GGGTCACTTCGGTTGACACT Chr7:26315488..26315507 60.01 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000636359 Chr7:26324720..26324806 TCCCTGGTTTGGACTACTGC Chr7:26324726..26324745 60.11 55
upstream ENSMUSE00000461821 Chr7:26323593..26323952 AAGGGAGTCTCGGCCTTTAG Chr7:26323820..26323839 59.84 55
upstream ENSMUSE00000460573 Chr7:26315490..26315774 AACCGAAGTGACCCATTCAG Chr7:26315504..26315523 59.97 50

*** Putative Vector Insertion (Chr 7: 26315489 - 26323953) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000462771 Chr7:26314769..26315023 GGGGATATAATCGGGGTGTC Chr7:26314974..26314993 60.23 55
downstream ENSMUSE00000199496 Chr7:26312760..26313035 CGCTGACTGGATTCGAGATT Chr7:26312774..26312793 60.36 50
downstream ENSMUSE00000199486 Chr7:26305833..26305950 ATGCCAGCCCTGCTATTAGA Chr7:26305839..26305858 59.83 50
downstream ENSMUSE00000598264 Chr7:26304982..26305034 TGTGAGATCTCGCTGGTCAC Chr7:26304988..26305007 59.99 55
downstream ENSMUSE00000636355 Chr7:26304178..26304209 No primer for this exon
downstream ENSMUSE00000636358 Chr7:26304178..26304209 No primer for this exon
downstream ENSMUSE00000598262 Chr7:26301062..26303204 CAGAGGGTGTGCCTTAGCTC Chr7:26301281..26301300 60.01 60
downstream ENSMUSE00000636354 Chr7:26301062..26303204 CAGAGGGTGTGCCTTAGCTC Chr7:26301281..26301300 60.01 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCCTCACTTTTAGCCTCCTG Chr7:26323932..26323953 60.03 52.38 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGACTGTCGTGACTGGGAAA Chr7:26320889..26320909 60.28 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000054385