Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6704
Trapped Gene
Rab5a (ENSMUSG00000017831)
Vector Insertion
Chr 17: 53623401 - 53636851
Public Clones BGC520 (baygenomics) XH206 (baygenomics) BGC512 (baygenomics)
Private Clones OST427098 (lexicon) OST381336 (lexicon)
Severity of mutation (?) Insertion after 25% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000364151 (Chr17:53623146..53623400 +)
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
ENSMUSE00000364151 (Chr17:53623146..53623400 +)
Downstram Exon
ENSMUSE00000542060 (Chr17:53636852..53637003 +)
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 ENSMUSE00000321375 Chr17:53618559..53618933 No primer for this exon
upstream ENSMUSE00000364151 Chr17:53623146..53623400 No primer for this exon

*** Putative Vector Insertion (Chr 17: 53623401 - 53636851) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000542060 Chr17:53636852..53637003 No primer for this exon
downstream ENSMUSE00000136476 Chr17:53637713..53637835 No primer for this exon
downstream ENSMUSE00000542056 Chr17:53639701..53639794 No primer for this exon
downstream ENSMUSE00000542054 Chr17:53645639..53646996 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GAGTACCATTGGGGGTGAGA Chr17:53635388..53635408 59.78 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AAATTTTCGTGACTGGGAAAAC Chr17:53635445..53635467 59.39 36.36 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000017831