Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI22344
Trapped Gene
Slc25a30 (ENSMUSG00000022003)
Vector Insertion
Chr 14: 76171341 - 76174801
Public Clones not available
Private Clones OST299643 (lexicon)
Severity of mutation (?) Insertion after 24% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000122869 (Chr14:76174802..76174949 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGGATAGGCCGAGAAGAAGG Chr14:76174823..76174842 59.81 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000122869 (Chr14:76174802..76174949 -)
Downstram Exon
ENSMUSE00000122865 (Chr14:76171246..76171340 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGGATAGGCCGAGAAGAAGG Chr14:76174823..76174842 59.81 55 GTTCCACAGCTAACCGCTTC Chr14:76171233..76171252 59.88 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000275811 Chr14:76186732..76186841 GCTCCGAGGAGTAGCAGGAG Chr14:76186747..76186766 61.59 65
upstream ENSMUSE00000275801 Chr14:76176803..76176933 ATGTCAGCCCTCAACTGGAA Chr14:76176847..76176866 60.66 50
upstream ENSMUSE00000122869 Chr14:76174802..76174949 AGGATAGGCCGAGAAGAAGG Chr14:76174823..76174842 59.81 55

*** Putative Vector Insertion (Chr 14: 76171341 - 76174801) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000122865 Chr14:76171246..76171340 GTTCCACAGCTAACCGCTTC Chr14:76171233..76171252 59.88 55
downstream ENSMUSE00000275770 Chr14:76169978..76170063 ACATTGACCAGCAGGGTTTC Chr14:76170020..76170039 59.97 50
downstream ENSMUSE00000275762 Chr14:76169373..76169468 CCTCCTGCTGGTAAATGCTC Chr14:76169371..76169390 59.84 55
downstream ENSMUSE00000122871 Chr14:76168623..76168747 TCAGGTGCTTCTTGGTGATG Chr14:76168644..76168663 59.83 50
downstream ENSMUSE00000122864 Chr14:76166710..76166848 GTACCCTTATAGCCCGCACA Chr14:76166707..76166726 59.98 55
downstream ENSMUSE00000122867 Chr14:76163136..76163216 No primer for this exon
downstream ENSMUSE00000275734 Chr14:76160878..76162497 CAGCCGCCTCTGTAGTTTTC Chr14:76161765..76161784 60.01 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AATAATCGCCTTGCAGCACA Chr14:76171732..76171752 61.69 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCAAGGAATGTTTATGCGTTT Chr14:76171766..76171787 59.14 38.1 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022003