Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32913
Trapped Gene
Tbc1d22b (ENSMUSG00000042203)
Vector Insertion
Chr 17: 29737187 - 29741788
Public Clones IST11444E5 (tigm) IST11444E3 (tigm) IST11444E3 (tigm) IST11444E5 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 92% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000234586 (Chr17:29737091..29737186 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCTTCTCGCACTTCCATCTC Chr17:29737104..29737123 60.1 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000234586 (Chr17:29737091..29737186 +)
Downstram Exon
ENSMUSE00000472519 (Chr17:29741789..29743751 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCTTCTCGCACTTCCATCTC Chr17:29737104..29737123 60.1 55 TGCTTCACCCTGGAAATAGG Chr17:29742977..29742996 60.07 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000548944 Chr17:29686747..29686939 CAGCAAGCAGTTCTGGAAGA Chr17:29686898..29686917 59.31 50
upstream ENSMUSE00000417039 Chr17:29701377..29701433 GGATCCTCGGCTCACAAAAA Chr17:29701413..29701432 62.41 50
upstream ENSMUSE00000417031 Chr17:29705337..29705644 ACTGCAGCCCAAGTCCTAGA Chr17:29705509..29705528 60.01 55
upstream ENSMUSE00000417027 Chr17:29707532..29707711 AATGACAGTCCGGGAGAAAA Chr17:29707641..29707660 59.53 45
upstream ENSMUSE00000417022 Chr17:29708610..29708680 CAGTGACCTGGAGACTGCTG Chr17:29708658..29708677 59.6 60
upstream ENSMUSE00000234820 Chr17:29709915..29710043 AATGAAGAGCACCACCAGGA Chr17:29710011..29710030 60.66 50
upstream ENSMUSE00000393319 Chr17:29712101..29712166 GAACCCTCTCATCCCCTTGT Chr17:29712124..29712143 60.31 55
upstream ENSMUSE00000234776 Chr17:29715774..29715888 TGTCGTCTTCCTCTCGGAAT Chr17:29715863..29715882 59.8 50
upstream ENSMUSE00000394496 Chr17:29731622..29731728 GGACGTGGAGAACTTTGACG Chr17:29731626..29731645 60.69 55
upstream ENSMUSE00000234625 Chr17:29732793..29732868 CCAGGGATCCAGAAGAAGGT Chr17:29732814..29732833 60.45 55
upstream ENSMUSE00000234607 Chr17:29736784..29736911 TTTCGGTGGATGAACAACCT Chr17:29736837..29736856 60.35 45
upstream ENSMUSE00000234586 Chr17:29737091..29737186 GCTTCTCGCACTTCCATCTC Chr17:29737104..29737123 60.1 55

*** Putative Vector Insertion (Chr 17: 29737187 - 29741788) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000472519 Chr17:29741789..29743751 TGCTTCACCCTGGAAATAGG Chr17:29742977..29742996 60.07 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATAATCGCCTTGCAGCACAT Chr17:29740237..29740257 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTGTAGCGTGACTGGGAAAA Chr17:29740232..29740252 59.32 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000042203