Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI654
Trapped Gene
AC113019.13 (ENSMUSG00000078669)
Vector Insertion
Chr 7: 82714918 - 82724387
Public Clones DC0594 (sanger)
Private Clones OST230036 (lexicon)
Severity of mutation (?) Insertion after 21% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000634165 (Chr7:82714770..82714917 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCACAAGTACCCGGAAAGTC Chr7:82714867..82714886 59.6 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000634165 (Chr7:82714770..82714917 +)
Downstram Exon
ENSMUSE00000634163 (Chr7:82724388..82724684 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCACAAGTACCCGGAAAGTC Chr7:82714867..82714886 59.6 55 CAGGTGTCTGAATGCCAGTG Chr7:82724638..82724657 60.31 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000634167 Chr7:82600443..82600634 CTCTTGGGCTCGGTTGAGAC Chr7:82600486..82600505 62.28 60
upstream ENSMUSE00000634166 Chr7:82679694..82679737 CCTGGGTCATGAAACTTAGTCC Chr7:82679697..82679718 59.86 50
upstream ENSMUSE00000634165 Chr7:82714770..82714917 GCACAAGTACCCGGAAAGTC Chr7:82714867..82714886 59.6 55

*** Putative Vector Insertion (Chr 7: 82714918 - 82724387) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000634163 Chr7:82724388..82724684 CAGGTGTCTGAATGCCAGTG Chr7:82724638..82724657 60.31 55
downstream ENSMUSE00000634162 Chr7:82731043..82731226 CCTTCCTTGTTGTGGATGCT Chr7:82731166..82731185 60.11 50
downstream ENSMUSE00000634161 Chr7:82747672..82748187 ATGGGGTTCACGATGTGACT Chr7:82747810..82747829 60.25 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ACGATTAGGCGGCTAAGACC Chr7:82714912..82714932 60.6 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACGATTAGGCGGCTAAGACC Chr7:82714912..82714932 60.6 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000078669