Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20095
Trapped Gene
Ptges (ENSMUSG00000050737)
Vector Insertion
Chr 2: 30748117 - 30748347
Public Clones IST10874B4 (tigm)
Private Clones OST373055 (lexicon) OST355894 (lexicon) OST286382 (lexicon) OST14968 (lexicon)
Severity of mutation (?) Insertion after 39% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000568154 (Chr2:30748118..30748346 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCTCGGCTTCGTGTACTCAT Chr2:30748285..30748304 60.28 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000568154 (Chr2:30748118..30748346 -)
Downstram Exon
ENSMUSE00000662812 (Chr2:30744995..30748346 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCTCGGCTTCGTGTACTCAT Chr2:30748285..30748304 60.28 55 CAGCCTAATGTTCAGCGACA Chr2:30746568..30746587 60.01 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000662813 Chr2:30758610..30758819 CTCACTCTCAGTCCCGGTGT Chr2:30758771..30758790 60.31 60
upstream ENSMUSE00000696302 Chr2:30757234..30757308 GAACTCCACTGGTGGCAAAG Chr2:30757282..30757301 60.69 55
upstream ENSMUSE00000401188 Chr2:30756387..30756469 No primer for this exon
upstream ENSMUSE00000568154 Chr2:30748118..30748346 CCTCGGCTTCGTGTACTCAT Chr2:30748285..30748304 60.28 55

*** Putative Vector Insertion (Chr 2: 30748117 - 30748347) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000696303 Chr2:30748086..30748115 AGCTGCTGGTCACAGATGGT Chr2:30748066..30748085 60.89 55
downstream ENSMUSE00000662812 Chr2:30744995..30748346 CAGCCTAATGTTCAGCGACA Chr2:30746568..30746587 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCTTTCCTCTTCCTCGGCTA Chr2:30748294..30748314 60.83 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCAACGACATGGAGACAATCT Chr2:30748316..30748337 60.13 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CCTTTCCTCTTCCTCGGCTA Chr2:30745294..30745314 60.83 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CGCAACGACATGGAGACAAT Chr2:30745318..30745338 62.07 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000050737