Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI1918
Trapped Gene
Tcfap4 (ENSMUSG00000005718)
Vector Insertion
Chr 16: 4549472 - 4551229
Public Clones AX0650 (sanger) IST10375C4 (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
ENSMUSE00000395356 (Chr16:4551230..4551400 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000395356 (Chr16:4551230..4551400 -)
Downstram Exon
ENSMUSE00000371658 (Chr16:4549331..4549471 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000410136 Chr16:4559428..4559854 No primer for this exon
upstream ENSMUSE00000127814 Chr16:4551935..4552100 No primer for this exon
upstream ENSMUSE00000127813 Chr16:4551746..4551844 No primer for this exon
upstream ENSMUSE00000395356 Chr16:4551230..4551400 No primer for this exon

*** Putative Vector Insertion (Chr 16: 4549472 - 4551229) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000371658 Chr16:4549331..4549471 No primer for this exon
downstream ENSMUSE00000400961 Chr16:4547255..4547410 No primer for this exon
downstream ENSMUSE00000370959 Chr16:4544664..4545747 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTGATGCCCAGAGTCAGAG Chr16:4551182..4551202 59.98 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGTGATGCCCAGAGTCAGAG Chr16:4551182..4551202 59.98 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CTCACCAACCTCTTGGGTTC Chr16:4551404..4551424 59.55 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CTCACCAACCTCTTGGGTTC Chr16:4551404..4551424 59.55 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000005718