Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI13590
Trapped Gene
Tnfrsf10b (ENSMUSG00000022074)
Vector Insertion
Chr 14: 70180381 - 70182038
Public Clones CMHD-GT_309G07-3 (cmhd) CMHD-GT_311F04-3 (cmhd) PST15354-NR (escells) PST224-1 (escells)
PST5759-NR (escells) PST2098-NR (escells)
Private Clones not available
Severity of mutation (?) Insertion after 71% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000412290 (Chr14:70180311..70180380 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAGTTGCTGGTTCCGGTAAA Chr14:70180341..70180360 59.61 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000412290 (Chr14:70180311..70180380 +)
Downstram Exon
ENSMUSE00000371745 (Chr14:70182039..70184210 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAGTTGCTGGTTCCGGTAAA Chr14:70180341..70180360 59.61 45 GTGTTTCGGCTTTGACCATT Chr14:70182158..70182177 59.98 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000263828 Chr14:70167336..70167598 ATAGCTTTGCGTTGCTGCTT Chr14:70167567..70167586 60.18 45
upstream ENSMUSE00000263817 Chr14:70170588..70170684 TAACCCAGCCCATAATCGTC Chr14:70170611..70170630 59.78 50
upstream ENSMUSE00000123392 Chr14:70173184..70173294 TGACTACACCAGCCATTCCA Chr14:70173230..70173249 60.11 50
upstream ENSMUSE00000123395 Chr14:70174893..70175001 AATACGGTGTGTCGATGCAA Chr14:70174928..70174947 59.99 45
upstream ENSMUSE00000123398 Chr14:70175879..70176075 ACTGACGGGGAAGAGGAACT Chr14:70175883..70175902 60.11 55
upstream ENSMUSE00000348805 Chr14:70177528..70177622 TGTGCATTCGTCTCTCTTGG Chr14:70177559..70177578 59.98 50
upstream ENSMUSE00000412290 Chr14:70180311..70180380 AAGTTGCTGGTTCCGGTAAA Chr14:70180341..70180360 59.61 45

*** Putative Vector Insertion (Chr 14: 70180381 - 70182038) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000371745 Chr14:70182039..70184210 GTGTTTCGGCTTTGACCATT Chr14:70182158..70182177 59.98 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AATAGCACAGAAGCCCAAGG Chr14:70180402..70180422 59.34 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AATAGCACAGAAGCCCAAGG Chr14:70180402..70180422 59.34 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022074