Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI4324
Trapped Gene
Lmnb1 (ENSMUSG00000024590)
Vector Insertion
Chr 18: 56909464 - 56912260
Public Clones AG0082 (sanger)
Private Clones OST470561 (lexicon) OST334988 (lexicon) OST23329 (lexicon)
Severity of mutation (?) Insertion after 98% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000492660 (Chr18:56909353..56909463 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGGAGGAGGAGGAGGAGGAG Chr18:56909402..56909421 61.24 65 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000492660 (Chr18:56909353..56909463 +)
Downstram Exon
ENSMUSE00000489632 (Chr18:56912261..56913076 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGGAGGAGGAGGAGGAGGAG Chr18:56909402..56909421 61.24 65 AGCTTGAGGAAGATCGACCA Chr18:56912338..56912357 59.95 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000625385 Chr18:56867467..56868128 AGCCCGAGAGGAAACAAAGT Chr18:56867514..56867533 60.25 50
upstream ENSMUSE00000143569 Chr18:56888055..56888211 GCCCTAGGGGACAAAAAGAG Chr18:56888149..56888168 60.07 55
upstream ENSMUSE00000143564 Chr18:56888941..56889066 GGATTTGGAGAATCGCTGTC Chr18:56889000..56889019 59.63 50
upstream ENSMUSE00000143567 Chr18:56890624..56890794 TGGGCGTCAGATTGAGTATG Chr18:56890677..56890696 59.67 50
upstream ENSMUSE00000143562 Chr18:56892866..56892991 No primer for this exon
upstream ENSMUSE00000143565 Chr18:56899475..56899695 AGATCAGGGACCAGATGCAG Chr18:56899578..56899597 60.22 55
upstream ENSMUSE00000143571 Chr18:56900341..56900566 GGAAAGCGGAAGAGAGTTGA Chr18:56900420..56900439 59.55 50
upstream ENSMUSE00000494265 Chr18:56902865..56902969 GGCTGGGAGATGATCAGAAA Chr18:56902880..56902899 60.16 50
upstream ENSMUSE00000491699 Chr18:56907631..56907750 CCCTCCAACTGACCTCATCT Chr18:56907663..56907682 59.1 55
upstream ENSMUSE00000492660 Chr18:56909353..56909463 AGGAGGAGGAGGAGGAGGAG Chr18:56909402..56909421 61.24 65

*** Putative Vector Insertion (Chr 18: 56909464 - 56912260) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000489632 Chr18:56912261..56913076 AGCTTGAGGAAGATCGACCA Chr18:56912338..56912357 59.95 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CACCAGCAGGTACGGTTTTC Chr18:56909456..56909476 60.55 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAAAAGCCTGTTTGGGTTTC Chr18:56909496..56909516 59.59 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024590