Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI27272
Trapped Gene
Ostb (ENSMUSG00000053862)
Vector Insertion
Chr 9: 65261822 - 65262973
Public Clones not available
Private Clones OST65266 (lexicon)
Severity of mutation (?) Insertion after 24% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000422571 (Chr9:65262974..65263094 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGCAGAAACATGGACCACA Chr9:65263055..65263074 59.68 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000422571 (Chr9:65262974..65263094 -)
Downstram Exon
ENSMUSE00000422553 (Chr9:65261731..65261821 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGCAGAAACATGGACCACA Chr9:65263055..65263074 59.68 50 CCTTCTCAGGAGGAACATGC Chr9:65261726..65261745 59.8 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000636032 Chr9:65270554..65270762 TCGGAAGAGTTCCTGTGAGG Chr9:65270623..65270642 60.38 55
upstream ENSMUSE00000422571 Chr9:65262974..65263094 GAGCAGAAACATGGACCACA Chr9:65263055..65263074 59.68 50

*** Putative Vector Insertion (Chr 9: 65261822 - 65262973) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000422553 Chr9:65261731..65261821 CCTTCTCAGGAGGAACATGC Chr9:65261726..65261745 59.8 55
downstream ENSMUSE00000422560 Chr9:65260521..65260808 TAAGTTAGGCGGCGTTATGG Chr9:65260535..65260554 60.11 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CATTCTCCTAATCGCCTTGC Chr9:65262910..65262930 59.81 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CGTGCAGAAGATGGTAGGTG Chr9:65262965..65262985 59.31 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000053862