Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33660
Trapped Gene
Lctl (ENSMUSG00000032401)
Vector Insertion
Chr 9: 63969488 - 63969912
Public Clones IST10006C3 (tigm) IST10006C3 (tigm) IST10006B3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000719836 (Chr9:63969413..63969487 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCTCCTGCATCTACCTCCTG Chr9:63969452..63969471 59.82 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000719836 (Chr9:63969413..63969487 +)
Downstram Exon
ENSMUSE00000422538 (Chr9:63969913..63970041 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCTCCTGCATCTACCTCCTG Chr9:63969452..63969471 59.82 60 CACTGAATGTGAGCCAATGC Chr9:63970036..63970055 60.27 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000356415 Chr9:63964954..63965107 GCCCAGGAGTCATTCAGAGA Chr9:63965032..63965051 60.35 55
upstream ENSMUSE00000344302 Chr9:63965422..63965655 CATGGAGGAAGGTGAGGTGT Chr9:63965520..63965539 59.96 55
upstream ENSMUSE00000377548 Chr9:63966587..63966750 GGCAGTTCTGCCTACCAGAC Chr9:63966604..63966623 59.87 60
upstream ENSMUSE00000219425 Chr9:63967235..63967322 TCAGCCACTACCGATTTTCC Chr9:63967266..63967285 60.07 50
upstream ENSMUSE00000219431 Chr9:63967546..63967655 AGCGATTTCATTGACGCTCT Chr9:63967581..63967600 59.99 45
upstream ENSMUSE00000719836 Chr9:63969413..63969487 CCTCCTGCATCTACCTCCTG Chr9:63969452..63969471 59.82 60

*** Putative Vector Insertion (Chr 9: 63969488 - 63969912) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000422538 Chr9:63969913..63970041 CACTGAATGTGAGCCAATGC Chr9:63970036..63970055 60.27 50
downstream ENSMUSE00000219433 Chr9:63970130..63970225 GCAGCCACATATAGGCCAGT Chr9:63970212..63970231 60.12 55
downstream ENSMUSE00000219426 Chr9:63974508..63974562 GCTGCGCCACGTGTTATTAT Chr9:63974555..63974574 61.05 50
downstream ENSMUSE00000219424 Chr9:63974645..63974806 CTCAATATCGTCGGGGTTGT Chr9:63974718..63974737 59.81 50
downstream ENSMUSE00000323214 Chr9:63979357..63979631 CTTGCTCGGCACTCTTTGTT Chr9:63979380..63979399 60.57 50
downstream ENSMUSE00000219441 Chr9:63980610..63980736 TATGGGAGGGTCACCGTACT Chr9:63980636..63980655 59.28 55
downstream ENSMUSE00000332982 Chr9:63980905..63981104 GCACAGAAGCCTTGGGATAG Chr9:63981057..63981076 59.84 55
downstream ENSMUSE00000406376 Chr9:63982659..63982722 GCTTTGAGACGCCAACTTTC Chr9:63982684..63982703 60 50
downstream ENSMUSE00000410971 Chr9:63985613..63985923 CTGTGGGGACCACAATCTCT Chr9:63985666..63985685 59.96 55
downstream ENSMUSE00000718716 Chr9:63985716..63985912 AGAGGAGATCCCGTCTCAGC Chr9:63985746..63985765 60.9 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTTCCAGCTTAATCGCCTTG Chr9:63969530..63969550 60.33 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTCCTAAGCAGGCATTTCCA Chr9:63969516..63969536 60.71 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032401