Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI12239
Trapped Gene
Rprd1b (ENSMUSG00000027651)
Vector Insertion
Chr 2: 157875950 - 157884387
Public Clones A047F07 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 97% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000316148 (Chr2:157875823..157875949 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAAGACGTGTCGCTGCTAGA Chr2:157875919..157875938 60.35 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000316148 (Chr2:157875823..157875949 +)
Downstram Exon
ENSMUSE00000404617 (Chr2:157884388..157884563 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAAGACGTGTCGCTGCTAGA Chr2:157875919..157875938 60.35 55 GGGTGTATTCCACCAGCATC Chr2:157884522..157884541 60.2 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000661331 Chr2:157854231..157854781 TAAAGCGCATTTTCGGTGTT Chr2:157854264..157854283 60.62 40
upstream ENSMUSE00000639455 Chr2:157854261..157854781 TAAAGCGCATTTTCGGTGTT Chr2:157854264..157854283 60.62 40
upstream ENSMUSE00000383532 Chr2:157854437..157854781 GCTGTTACTGCGGAGACCTG Chr2:157854472..157854491 61 60
upstream ENSMUSE00000171485 Chr2:157861732..157861861 TGTGCTTGTGGATGCTTTTT Chr2:157861829..157861848 59.32 40
upstream ENSMUSE00000680612 Chr2:157861732..157861861 TGTGCTTGTGGATGCTTTTT Chr2:157861829..157861848 59.32 40
upstream ENSMUSE00000333625 Chr2:157869142..157869275 TCTGGCAAGAACGAAGTGTG Chr2:157869189..157869208 60.03 50
upstream ENSMUSE00000680611 Chr2:157869142..157869275 TCTGGCAAGAACGAAGTGTG Chr2:157869189..157869208 60.03 50
upstream ENSMUSE00000386995 Chr2:157873652..157873764 TGGAAGCTACTCTCCCCAAG Chr2:157873722..157873741 59.42 55
upstream ENSMUSE00000661330 Chr2:157873655..157873764 TGGAAGCTACTCTCCCCAAG Chr2:157873722..157873741 59.42 55
upstream ENSMUSE00000316148 Chr2:157875823..157875949 CAAGACGTGTCGCTGCTAGA Chr2:157875919..157875938 60.35 55

*** Putative Vector Insertion (Chr 2: 157875950 - 157884387) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000404617 Chr2:157884388..157884563 GGGTGTATTCCACCAGCATC Chr2:157884522..157884541 60.2 55
downstream ENSMUSE00000680609 Chr2:157896192..157896245 CATTGACAAGCATGCACAGA Chr2:157896221..157896240 59.41 45
downstream ENSMUSE00000639454 Chr2:157896297..157896350 TGGAGCTGCTGAGTAAGACG Chr2:157896319..157896338 59.34 55
downstream ENSMUSE00000680613 Chr2:157896297..157896392 CAGGGCAGCTACCACATTTC Chr2:157896371..157896390 60.66 55
downstream ENSMUSE00000343617 Chr2:157900666..157903931 TAGGCAACAGCGAAAGGTCT Chr2:157900759..157900778 60.01 50
downstream ENSMUSE00000661329 Chr2:157900666..157903943 TAGGCAACAGCGAAAGGTCT Chr2:157900759..157900778 60.01 50
downstream ENSMUSE00000680614 Chr2:157956875..157956894 TCATGCCTCAGTTTCTGCAT Chr2:157956897..157956916 59.4 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CGTGTCGCTGCTAGAGAAAA Chr2:157881925..157881945 59.37 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAGTCGTGACTGGGAAAACC Chr2:157881997..157882017 59.56 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027651