Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI4310
Trapped Gene
Gabpb1 (ENSMUSG00000027361)
Vector Insertion
Chr 2: 126464916 - 126465081
Public Clones AG0132 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 65% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000322266 (Chr2:126464965..126465080 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TCATCAGTGAAGAGCCACCA Chr2:126465022..126465041 60.4 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000322266 (Chr2:126464965..126465080 -)
Downstram Exon
ENSMUSE00000661640 (Chr2:126464917..126465080 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TCATCAGTGAAGAGCCACCA Chr2:126465022..126465041 60.4 50 CCCGGCTCTCAATTATTTCC Chr2:126464963..126464982 60.76 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000683920 Chr2:126501770..126502073 TGAAGAGGACAGGCATCCTT Chr2:126501823..126501842 59.8 50
upstream ENSMUSE00000398603 Chr2:126501092..126501304 GACTTTTCGGTGGGGATTTT Chr2:126501250..126501269 60.17 45
upstream ENSMUSE00000597120 Chr2:126501092..126501191 No primer for this exon
upstream ENSMUSE00000661644 Chr2:126501092..126501280 GACTTTTCGGTGGGGATTTT Chr2:126501250..126501269 60.17 45
upstream ENSMUSE00000661647 Chr2:126501092..126501328 GACTTTTCGGTGGGGATTTT Chr2:126501250..126501269 60.17 45
upstream ENSMUSE00000661643 Chr2:126488727..126488824 TCCTACCTGCTGACCAGCTC Chr2:126488799..126488818 60.56 60
upstream ENSMUSE00000641451 Chr2:126484206..126484313 TGAAGTTCGCATTTTGATGG Chr2:126484235..126484254 59.66 40
upstream ENSMUSE00000718562 Chr2:126484206..126484313 TGAAGTTCGCATTTTGATGG Chr2:126484235..126484254 59.66 40
upstream ENSMUSE00000168730 Chr2:126479291..126479458 CCGAGCCGGTGTAAGTAGAG Chr2:126479374..126479393 59.89 60
upstream ENSMUSE00000168732 Chr2:126477969..126478163 GTGCTGACGTCAATGCAAAG Chr2:126478140..126478159 60.46 50
upstream ENSMUSE00000168727 Chr2:126476245..126476356 GACAATACACGCTGCCACAC Chr2:126476287..126476306 60.19 55
upstream ENSMUSE00000168731 Chr2:126475804..126475917 CCCCATTGTCCAATTCTTCA Chr2:126475814..126475833 60.69 45
upstream ENSMUSE00000597119 Chr2:126475804..126475917 GCCCCATTGTCCAATTCTTC Chr2:126475815..126475834 61.59 50
upstream ENSMUSE00000641435 Chr2:126473484..126475917 GGGCTACCTCCTTGGGTAAG Chr2:126475195..126475214 59.96 60
upstream ENSMUSE00000322272 Chr2:126472299..126472484 GAGGAAGTGGTTACCGCAGA Chr2:126472454..126472473 60.26 55
upstream ENSMUSE00000597113 Chr2:126472299..126472484 GAGGAAGTGGTTACCGCAGA Chr2:126472454..126472473 60.26 55
upstream ENSMUSE00000661646 Chr2:126472299..126472481 GAGGAAGTGGTTACCGCAGA Chr2:126472454..126472473 60.26 55
upstream ENSMUSE00000322266 Chr2:126464965..126465080 TCATCAGTGAAGAGCCACCA Chr2:126465022..126465041 60.4 50
upstream ENSMUSE00000597117 Chr2:126464965..126465080 CATCAGTGAAGAGCCACCAG Chr2:126465021..126465040 59.42 55
upstream ENSMUSE00000661640 Chr2:126464917..126465080 ATACCCGGTGTGTTTTGCTG Chr2:126464933..126464952 60.81 50

*** Putative Vector Insertion (Chr 2: 126464916 - 126465081) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000683919 Chr2:126464372..126465080 TTGAGGGCTTGCCATATGTT Chr2:126464752..126464771 60.47 45
downstream ENSMUSE00000661642 Chr2:126454651..126456118 ATGGTCGCCAGTAAAACCAG Chr2:126455211..126455230 59.99 50
downstream ENSMUSE00000597112 Chr2:126454649..126456118 ATGGTCGCCAGTAAAACCAG Chr2:126455211..126455230 59.99 50
downstream ENSMUSE00000358516 Chr2:126453178..126456118 ATGGTCGCCAGTAAAACCAG Chr2:126455211..126455230 59.99 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTAACCTGTGCCTCCCTTT Chr2:126465087..126465107 59.59 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGTAACCTGTGCCTCCCTTT Chr2:126465087..126465107 59.59 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027361