Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI28930
Trapped Gene
Ube2l6 (ENSMUSG00000027078)
Vector Insertion
Chr 2: 84643678 - 84646493
Public Clones IST11826B11 (tigm) IST11826B11 (tigm) IST11893H4 (tigm) IST11696H4 (tigm)
IST10107G8 (tigm)
Private Clones OST15763 (lexicon)
Severity of mutation (?) Insertion after 27% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000688579 (Chr2:84643585..84643677 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGATCTGCCTGTTTGTCCTC Chr2:84643639..84643658 59.66 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000688579 (Chr2:84643585..84643677 +)
Downstram Exon
ENSMUSE00000165867 (Chr2:84646494..84646680 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGATCTGCCTGTTTGTCCTC Chr2:84643639..84643658 59.66 55 GTGGGAGGCTTGAATGGATA Chr2:84646573..84646592 59.89 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000389304 Chr2:84638996..84639089 AGTTCAGAGAGCGGTGTCCT Chr2:84639007..84639026 59.05 55
upstream ENSMUSE00000721893 Chr2:84638996..84639089 AGTTCAGAGAGCGGTGTCCT Chr2:84639007..84639026 59.05 55
upstream ENSMUSE00000235673 Chr2:84642936..84643031 CGCCAACTGTCTAGTGACGA Chr2:84642975..84642994 60.05 55
upstream ENSMUSE00000688579 Chr2:84643585..84643677 GGATCTGCCTGTTTGTCCTC Chr2:84643639..84643658 59.66 55

*** Putative Vector Insertion (Chr 2: 84643678 - 84646493) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000165867 Chr2:84646494..84646680 GTGGGAGGCTTGAATGGATA Chr2:84646573..84646592 59.89 50
downstream ENSMUSE00000235664 Chr2:84649162..84650333 GCACAGGCTCTTCCAGATTC Chr2:84649218..84649237 59.96 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr2:84643729..84643749 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector

Come back to gene ENSMUSG00000027078