Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI879
Trapped Gene
Cog8 (ENSMUSG00000031916)
Vector Insertion
Chr 8: 109574211 - 109576112
Public Clones CA0053 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 74% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000214632 (Chr8:109576113..109576940 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCTAAGCCAGCTGATCCAAC Chr8:109576883..109576902 59.99 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000214632 (Chr8:109576113..109576940 -)
Downstram Exon
ENSMUSE00000364072 (Chr8:109574042..109574210 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCTAAGCCAGCTGATCCAAC Chr8:109576883..109576902 59.99 55 GGACTTGAAGACAGCGGTTC Chr8:109574053..109574072 59.85 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000346388 Chr8:109580161..109580584 CGCCAACTACAAGACGTTCA Chr8:109580267..109580286 59.9 50
upstream ENSMUSE00000611318 Chr8:109580161..109580583 CGCCAACTACAAGACGTTCA Chr8:109580267..109580286 59.9 50
upstream ENSMUSE00000214631 Chr8:109577922..109578129 TGACTCTAAACCGGCACACA Chr8:109578057..109578076 60.3 50
upstream ENSMUSE00000214632 Chr8:109576113..109576940 GCTAAGCCAGCTGATCCAAC Chr8:109576883..109576902 59.99 55

*** Putative Vector Insertion (Chr 8: 109574211 - 109576112) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000364072 Chr8:109574042..109574210 GGACTTGAAGACAGCGGTTC Chr8:109574053..109574072 59.85 55
downstream ENSMUSE00000414609 Chr8:109572683..109573030 GTCCAAGGTTTCCGTGCTTA Chr8:109572967..109572986 60.11 50
downstream ENSMUSE00000580190 Chr8:109572609..109573030 GTCCAAGGTTTCCGTGCTTA Chr8:109572967..109572986 60.11 50
downstream ENSMUSE00000335882 Chr8:109570195..109571171 ATGATTCGAGCTGTCCAACC Chr8:109571089..109571108 60.08 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATGTGACTGGGACCTTGGAA Chr8:109576126..109576146 60.36 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ATGTGACTGGGACCTTGGAA Chr8:109576126..109576146 60.36 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031916