Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI35641
Trapped Gene
Golga3 (ENSMUSG00000029502)
Vector Insertion
Chr 5: 110645993 - 110646829
Public Clones IST14591G11 (tigm) IST12981D12 (tigm) IST14640D7 (tigm) IST14935H3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 83% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000299024 (Chr5:110645853..110645992 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAGAATTGCAGGCCAAAGAA Chr5:110645932..110645951 59.82 40 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000299024 (Chr5:110645853..110645992 +)
Downstram Exon
ENSMUSE00000189591 (Chr5:110646830..110646962 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAGAATTGCAGGCCAAAGAA Chr5:110645932..110645951 59.82 40 GCCAGCTCAGTCTGGAATTG Chr5:110646886..110646905 60.94 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000445555 Chr5:110605720..110605941 TCTGTCCGGAGATCCATACC Chr5:110605755..110605774 59.89 55
upstream ENSMUSE00000409750 Chr5:110610622..110610932 GAGCATCAGCCAAACAGGAT Chr5:110610804..110610823 60.23 50
upstream ENSMUSE00000713219 Chr5:110610622..110610932 GAGCATCAGCCAAACAGGAT Chr5:110610804..110610823 60.23 50
upstream ENSMUSE00000540529 Chr5:110613357..110613626 TCCCAGACCTTCAGTTGTCC Chr5:110613444..110613463 60.09 55
upstream ENSMUSE00000691944 Chr5:110613357..110613506 AGTTGTCCCTTGATCCCACA Chr5:110613456..110613475 60.36 50
upstream ENSMUSE00000299106 Chr5:110614805..110614917 GTACCGAGTTAAGCGCCATC Chr5:110614890..110614909 59.74 55
upstream ENSMUSE00000299102 Chr5:110616360..110617006 GGAGCCGTAGAGACAGCATC Chr5:110616979..110616998 59.98 60
upstream ENSMUSE00000299095 Chr5:110617682..110617793 GGCTGAACCACAGGATGAAT Chr5:110617703..110617722 59.93 50
upstream ENSMUSE00000299090 Chr5:110618625..110618931 ACAAGTGGCAGAGGAGCAGT Chr5:110618849..110618868 60.06 55
upstream ENSMUSE00000299085 Chr5:110621926..110622128 GCAGATGACAGCGCTACAGA Chr5:110621942..110621961 60.32 55
upstream ENSMUSE00000299078 Chr5:110622638..110622775 AGCATCGATCCATCAAGGAG Chr5:110622720..110622739 60.18 50
upstream ENSMUSE00000299072 Chr5:110630850..110631011 AAGGTGAACGGGAGCAGTTA Chr5:110630944..110630963 59.73 50
upstream ENSMUSE00000189582 Chr5:110631588..110631956 GCTCTACAATCGGCCAAGAG Chr5:110631828..110631847 59.98 55
upstream ENSMUSE00000189585 Chr5:110632739..110632816 CAGCCACGTTGAGAAAACAG Chr5:110632761..110632780 59.49 50
upstream ENSMUSE00000189595 Chr5:110633799..110634062 CTCACGACTGCAGAAGGACA Chr5:110633954..110633973 60.18 55
upstream ENSMUSE00000189600 Chr5:110634796..110634890 CTTACCGAGGCCAATGAGAC Chr5:110634829..110634848 59.69 55
upstream ENSMUSE00000189592 Chr5:110636575..110636791 GGAGGCTACAGATGCAGAGC Chr5:110636719..110636738 60.13 60
upstream ENSMUSE00000189586 Chr5:110637459..110637602 AGATTCGTGCTCTGGAGGTG Chr5:110637463..110637482 60.41 55
upstream ENSMUSE00000189597 Chr5:110638794..110638991 GAGCACAACAGCATTCTGGA Chr5:110638923..110638942 59.99 50
upstream ENSMUSE00000299028 Chr5:110641250..110641366 GATCGCCAGATGAAGCAACT Chr5:110641283..110641302 60.37 50
upstream ENSMUSE00000299024 Chr5:110645853..110645992 AAGAATTGCAGGCCAAAGAA Chr5:110645932..110645951 59.82 40

*** Putative Vector Insertion (Chr 5: 110645993 - 110646829) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000189591 Chr5:110646830..110646962 GCCAGCTCAGTCTGGAATTG Chr5:110646886..110646905 60.94 55
downstream ENSMUSE00000189596 Chr5:110647113..110647235 TGTGTCCCCTCCAGTTCTTT Chr5:110647225..110647244 59.55 50
downstream ENSMUSE00000189588 Chr5:110649801..110649962 ATCCAGCTTGAGCTGTTGGT Chr5:110649944..110649963 59.87 50
downstream ENSMUSE00000189601 Chr5:110650643..110650806 GGACAGGGCAATCTGGTATC Chr5:110650718..110650737 59.37 55
downstream ENSMUSE00000356242 Chr5:110651927..110652172 No primer for this exon
downstream ENSMUSE00000691943 Chr5:110651927..110655489 AGAGTCCTCTGAACCGCAAA Chr5:110654370..110654389 59.99 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGTGGATGAGCTCCAGTGAG Chr5:110645979..110645999 60.83 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGTGGATGAGCTCCAGTGAG Chr5:110645979..110645999 60.83 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000029502