Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI8196
Trapped Gene
Rras2 (ENSMUSG00000055723)
Vector Insertion
Chr 7: 121193934 - 121202053
Public Clones RRH305 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 67% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000527807 (Chr7:121202054..121202162 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGCTGACCTGGACCATCAGA Chr7:121202059..121202078 60.83 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000527807 (Chr7:121202054..121202162 -)
Downstram Exon
ENSMUSE00000436813 (Chr7:121193815..121193933 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGCTGACCTGGACCATCAGA Chr7:121202059..121202078 60.83 55 GAGCTGCCTTGCTAACTGCT Chr7:121193873..121193892 59.93 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000588859 Chr7:121260910..121261295 GCTCACCATCCAGTTCATCC Chr7:121260912..121260931 60.48 55
upstream ENSMUSE00000527809 Chr7:121203846..121203933 CGGACTATGATCCAACCATTG Chr7:121203900..121203920 60.2 47.62
upstream ENSMUSE00000527808 Chr7:121202441..121202543 TACAGCCGGACAAGAGGAGT Chr7:121202517..121202536 59.87 55
upstream ENSMUSE00000527807 Chr7:121202054..121202162 AGCTGACCTGGACCATCAGA Chr7:121202059..121202078 60.83 55

*** Putative Vector Insertion (Chr 7: 121193934 - 121202053) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000436813 Chr7:121193815..121193933 GAGCTGCCTTGCTAACTGCT Chr7:121193873..121193892 59.93 55
downstream ENSMUSE00000504249 Chr7:121190302..121191765 GGGGACACTGAACGTGACTT Chr7:121190534..121190553 60.01 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGGAGGGCAAGTCAGTATGT Chr7:121199037..121199057 59.02 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGGAGGGCAAGTCAGTATGT Chr7:121199037..121199057 59.02 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000055723