Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI12960
Trapped Gene
Rab25 (ENSMUSG00000008601)
Vector Insertion
Chr 3: 88349179 - 88351954
Public Clones CMHD-GT_351E12-3 (cmhd) IST14254C4 (tigm) IST10613F9 (tigm) IST13723B4 (tigm)
IST12467B5 (tigm) IST12467A3 (tigm) IST13731D4 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 7% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000291599 (Chr3:88351955..88352186 -)
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
ENSMUSE00000291599 (Chr3:88351955..88352186 -)
Downstram Exon
ENSMUSE00000567123 (Chr3:88348983..88349178 -)
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 ENSMUSE00000291599 Chr3:88351955..88352186 No primer for this exon

*** Putative Vector Insertion (Chr 3: 88349179 - 88351954) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000567123 Chr3:88348983..88349178 No primer for this exon
downstream ENSMUSE00000567122 Chr3:88347242..88347435 No primer for this exon
downstream ENSMUSE00000175608 Chr3:88346623..88346703 No primer for this exon
downstream ENSMUSE00000291563 Chr3:88345958..88346292 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCTCTCAGCTCCCACAGAGT Chr3:88349186..88349206 59.58 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCTCTCAGCTCCCACAGAGT Chr3:88349186..88349206 59.58 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000008601